Growth promoting peptides and uses thereof

ABSTRACT

A natural peptide comprising a cellular growth promoting fragment of a protein selected from SEQUENCE ID NO&#39;s: 1 to 13, and a composition comprising a plurality of growth promoting peptides, is described. Also disclosed is the use of the peptides and compositions in prevention of ageing of human skin, treatment of diseases or conditions characterised by damaged epithelial cells or tissue such as colon cancer and peripheral inflammatory disorders, and wound treatment. Specific pea and rice protein derived peptides are described in SEQUENCE ID NO&#39;S 15 to 505, and 546 to 704.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has beensubmitted electronically in ASCII format and is hereby incorporated byreference in its entirety. Said ASCII copy, created on Oct. 30, 2019, isnamed 048262-091340-US_SL.txt and is 420,014 bytes in size.

BACKGROUND TO THE INVENTION

The growing will of maintaining a youthful appearance is leading to moreand more research of new dermatological procedures for treatment of skinaging, especially when people keeps living longer and healthier. Withage, structural and functional changes are be observed in human skin(Calleja-Agius J. et al. 2013) and many factors are responsible for themlike environmental factors such as UV radiation from sunlight orinternal factors such as changes in hormones due menopause (Affinito P.et al. 1999).

The alterations of connective tissue in the dermis and epidermis,especially the reduction of the extracellular matrix is highlyresponsible for skin wrinkling and sagging since they induce importantchanges in its mechanical properties. Furthermore, metabolism andsynthesis of the extra-cellular matrix are affected by the ageingprocess through enzymatic activities.

There are mainly four solutions to fight against ageing process at themoment:

Diet and hormones management: fighting the ageing process by managingour diet by using supplements. This method is still controverted at themoment. Some studies suggest that antioxidant supplements like Vitamin Cor lipoic acid for example could have anti-ageing properties.

Hormone treatment: a risky and controverted solution for anti-ageingpurpose. One have to be really careful with anything related to hormonessince it can have a broad spectrum of adverse effects. This solutionisn't widely used and most of the studies are still on the animal stage.

Surgical anti-aging solutions: they are very effective on the short termbut they are costly and can require long period of healing. Like inevery surgical operation, there are risks but also side effects. Forexample, during surgical anti-aging solutions like eyelifts andfacelifts, where an incision is made at the hairline, there are somerisks of bruising, swelling, drooping eyelids, and secondary infectionscan occur.

Recently, there has been an increasing enthusiasm on minimally invasivetreatments and techniques designed to deal with problems like wrinkles,volume loss and other skin damages. The most common topical anti-ageingsolutions are creams and serums. Their active ingredients can be dividedin several families:

Moisturiser ingredients will keep the skin hydrated. It's mostly bigpolar molecules that will make bonds with water like glycosaminoglycan(hyaluronic acid for instance).

Collagen and other extra-cellular matrix related ingredients willcontribute to maintain a well organised skin structure by stimulatingbiosynthesis if they can trigger the right receptors.

Cell proliferation ingredients are important as well since they help toregenerate skin cells which will synthesise the component the skin needslike extra-cellular matrix and growth factors.

It is an object of the invention to provide an alternative minimallyinvasive treatment of skin ageing.

STATEMENTS OF INVENTION

The pea genome codes for over 70,000 different proteins. The Applicanthas identified six of these proteins, each of which contains one or morepeptides capable of promoting cellular growth and/or proliferation(hereafter “growth promoting peptide” or “growth promoting fragment”).Likewise, out of the more than 60,000 proteins encoded by the ricegenome, the Applicant has identified seven proteins, each of whichcontains one or more peptides that are bioactive, typically capable ofpromoting cellular growth and/or proliferation. Cellular growthpromoting fragments of the fourteen identified proteins have been shownto have an effect on elastin and collagen production and cellproliferation (FIGS. 1 to 109). The specific plant proteins from whichthe natural peptides are derived are provided in SEQ ID NO: 1-14 and 705to 716. The specific pea proteins from which the peptides are derivedinclude SEQ ID NO: 1-2 and 7-10, and the specific rice proteins fromwhich the peptides are derived include SEQ ID NO: 3-6 and 11-13.Homologs of these proteins are described in SEQ ID NO: 525 to 564. Thespecific peptides initially identified in the pea proteins are shown inSEQ ID NO's: 15-215 and 283-340. The specific peptides initiallyidentified in the rice proteins are shown in SEQ ID NO's: 216-282 and341-412. Additional peptides identified in the pea and rice proteinsdisclosed herein, and variants thereof, are provided in SEQ ID NO'S 418to 515 and 546 to 704 and 717-775.

In a first aspect, the invention provides a peptide, typically 5 to 50amino acids in length, and comprising (a) a fragment of a pea or riceprotein, for example the pea and rice protein disclosed herein such asone selected from SEQ ID NO's: 1 to 14 and 705 to 716 and 717-732, or ahomolog thereof, or (b) a variant of the fragment, or (c) a fragment ofthe peptide (hereafter “peptide of the invention”). In one embodimentthe peptide is bioactive. In one embodiment, the peptide has cellulargrowth or proliferation promotion activity.

In one embodiment, the peptide of the invention comprises a sequenceselected from SEQ ID NO: 15-505 and 546-704 and 717-775.

In one embodiment, the peptide of the invention consists essentially ofa sequence selected from SEQ ID NO: 15-505 and 546-704 and 717-775.

In one embodiment, the peptide of the invention consists of about 3-50amino acids. In one embodiment, the peptide of the invention consists ofabout 4-50 amino acids. In one embodiment, the peptide of the inventionconsists of about 5-50 amino acids. In one embodiment, the peptide ofthe invention consists of about 6-50 amino acids. In one embodiment, thepeptide of the invention consists of about 7-50 amino acids.

In one embodiment, the fragment has 8 to 37 amino acids. In oneembodiment, the fragment has a charge of between −10 and +4.

Preferably, the c-terminal amino acid is not cysteine (C) or methionine(M).

Preferably, the n-terminal amino acid is not cysteine (C), histidine(H), or proline (P).

Preferably, the c-terminal domain of the fragment does not containcysteine (C).

Preferably, the n-terminal domain of the fragment does not containcysteine (C).

Preferably, the fragment does not contain cysteine (C).

Preferably, the peptide does not contain cysteine (C).

In one embodiment of the invention, the peptide comprises a sequenceselected from SEQ ID NO: 15 to 505.

In one embodiment of the invention, the peptide consists essentially ofa sequence selected from SEQ ID NO: 15 to 505.

Preferably, the fragment is selected from SEQ ID NO's: 15-215 and283-340, or a bioactive variant of the fragment.

Preferably, the fragment is selected from SEQ ID NO's: 216-282 and341-412, or a bioactive variant of the fragment.

Preferably, the peptide consists of a fragment selected from SEQ IDNO's: 15 to 417, or a bioactive variant of the fragment.

Preferably, the peptide consists of a sequence selected from SEQ IDNO's: 15 to 417.

Preferably, the fragment is selected from SEQ ID NO's:413-417, or abioactive variant of the fragment.

In one embodiment, the peptide comprises or consists of a fragmentselected from SEQ ID NO'S: 246, 283, 284, 245 and 42, or a bioactivevariant of the fragment.

In one embodiment, the peptide comprises a fragment selected from SEQ IDNO'S: 247 to 268.

In one embodiment, the peptide of the invention is modified. In oneembodiment the peptide is modified with a protecting group. In oneembodiment, the peptide is modified to increase its lipophilicity. Innone embodiment, the peptide is modified to increase its half-life. Inone embodiment, an N or C-terminal amino acid of the peptide ismodified. In one embodiment, the N or C-terminal amino acid of thepeptide is modified with a protecting group.

[SEQ ID NO: 1 (Pea Protein 1—P13918)]

Preferably, the peptide comprises a bioactive fragment of the protein ofSEQ ID NO: 1 or a homolog thereof, or a bioactive variant of thefragment.

Preferably, the peptide comprises a bioactive fragment selected from SEQID NO'S: 15 to 90, or a bioactive variant of the fragment.

The invention also provides a composition comprising at least 1,preferably at least 2, preferably at least 3, preferably at least 4,preferably at least 5, preferably at least 6, preferably at least 7,preferably at least 8, preferably at least 9, or preferably at least 10bioactive peptides of the invention, each of which typically comprises adifferent a bioactive fragment of SEQ ID NO: 1 or a homolog thereof.Preferably, the composition comprises a first bioactive peptidecomprising a first a bioactive fragment selected from SEQ ID NO: 15 to90 (or a bioactive variant of the fragment), and a second bioactivepeptide comprising a second a bioactive fragment selected from SEQ IDNO: 15-90 (or a bioactive variant of the fragment).

In one embodiment, the bioactive peptide, variant or fragment is acellular growth or proliferation promoting peptide, variant or fragment,respectively.

Homologs of Pea Protein 1 (SEQ ID NO: 1) include Vicia fabia, Cicerarietinum and Lens culinaris homologs (SEQ ID NO: 525 to 527).

[SEQ ID NO: 2 (Pea Protein 2—Q9M3X6)]

Preferably, the peptide comprises a bioactive fragment of the protein ofSEQ ID NO: 2, or a homolog thereof, or a bioactive variant of thefragment.

Preferably, the peptide comprises a bioactive fragment selected from SEQID NO'S: 91 to 215, or a bioactive variant of the fragment.

The invention also provides a composition comprising at least 1,preferably at least 2, preferably at least 3, preferably at least 4,preferably at least 5, preferably at least 6, preferably at least 7,preferably at least 8, preferably at least 9, or preferably at least 10bioactive peptides of the invention, each of which comprises a differenta bioactive fragment of SEQ ID NO: 2 or a homolog thereof. Preferably,the composition comprises a first bioactive peptide comprising abioactive fragment selected from SEQ ID NO 91-215, and a secondbioactive peptide comprising a bioactive fragment selected from SEQ IDNO 91-215.

In one embodiment, the bioactive peptide, variant or fragment is acellular growth or proliferation promoting peptide, variant or fragment,respectively.

Homologs of Pea Protein 2 (SEQ ID NO: 2) include Pisum abyssinicum,Lathyrus annuus, and Vicia villosa (SEQ ID NOS 528 to 530).

[SEQ ID NO: 3 (Rice Protein 1—QODEV5)]

Preferably, the peptide comprises a bioactive fragment of the protein ofSEQ ID NO: 3, or a homolog thereof, or a bioactive variant of thefragment.

Preferably, the peptide comprises a bioactive fragment selected from SEQID NO'S: 216-244, or a bioactive variant of the fragment.

The invention also provides a composition comprising at least 1,preferably at least 2, preferably at least 3, preferably at least 4,preferably at least 5, preferably at least 6, preferably at least 7,preferably at least 8, preferably at least 9, or preferably at least 10bioactive peptides of the invention, each of which comprises a differentbioactive fragment of SEQ ID NO: 3 or a homolog thereof. Preferably, thecomposition comprises a first bioactive peptide comprising a bioactivefragment selected from SEQ ID NO 216-244, and a second bioactive peptidecomprising a bioactive fragment selected from SEQ ID NO 216-244.

In one embodiment, the bioactive peptide, variant or fragment is acellular growth or proliferation promoting peptide, variant or fragment,respectively.

Homologs of Rice Protein 1 (SEQ ID NO: 15) include Oryza rufipogon,Oryza officinalis, Hordeum vulgare subsp. vulgare (SEQ ID NO: 531 to533).

[SEQ ID NO: 4 (Rice Protein 2—P14323)]

Preferably, the peptide comprises a bioactive fragment of the protein ofSEQ ID NO: 4, or a homolog thereof, or a bioactive variant of thefragment.

Preferably, the peptide comprises a bioactive fragment selected from SEQID NO'S: 245-246, or a bioactive variant of the fragment.

The invention also provides a composition comprising at least one ormore peptides of the invention that comprise different bioactivefragments of SEQ ID NO: 4 or a homolog thereof. Preferably, thecomposition comprises a first peptide comprising the bioactive fragmentSEQ ID NO: 245 and a second peptide comprising the bioactive fragmentSEQ ID NO: 246.

In one embodiment, the bioactive peptide, variant or fragment is acellular growth or proliferation promoting peptide, variant or fragment,respectively.

Homologs of Rice Protein 2 (SEQ ID NO: 4) include Oryza brachyantha, andZizania latifolia (SEQ ID NO: 534 to 536).

[SEQ ID NO: 5 (Rice Protein 3—P29835)]

Preferably, the peptide comprises a bioactive fragment of the protein ofSEQ ID NO: 5, or a homolog thereof, or a bioactive variant of thefragment.

Preferably, the peptide comprises a bioactive fragment selected from SEQID NO'S: 247-268, or a bioactive variant of the fragment.

The invention also provides a composition comprising at least 1,preferably at least 2, preferably at least 3, preferably at least 4,preferably at least 5, preferably at least 6, preferably at least 7,preferably at least 8, preferably at least 9, or preferably at least 10bioactive peptides of the invention, each of which comprises a differentbioactive fragment of SEQ ID NO: 5 or a homolog thereof. Preferably, thecomposition comprises a first bioactive peptide comprising a bioactivefragment selected from SEQ ID NO 247-268, and a second bioactive peptidecomprising a bioactive fragment selected from SEQ ID NO 247-268.

In one embodiment, the bioactive peptide, variant or fragment is acellular growth or proliferation promoting peptide, variant or fragment,respectively.

Homologs of Rice Protein 3 (SEQ ID NO: 5) include Zea Mays, Sorghumbicolor and Setaria italica (SEQ ID NOS 537 to 539).

[SEQ ID NO: 6 (Rice Protein 4—P14614)]

Preferably, the peptide comprises a bioactive fragment of the protein ofSEQ ID NO: 6 or a homolog thereof, or a bioactive variant of thefragment.

Preferably, the peptide comprises a bioactive fragment selected from SEQID NO'S: 269-282, or a bioactive variant of the fragment.

The invention also provides a composition comprising at least 1,preferably at least 2, preferably at least 3, preferably at least 4,preferably at least 5, preferably at least 6, preferably at least 7,preferably at least 8, preferably at least 9, or preferably at least 10bioactive peptides of the invention, each of which comprises a differentbioactive fragment of SEQ ID NO: 6 or a homolog thereof. Preferably, thecomposition comprises a first bioactive peptide comprising a bioactivefragment selected from SEQ ID NO 269-282, and a second bioactive peptidecomprising a bioactive fragment selected from SEQ ID NO 269-282.

In one embodiment, the bioactive peptide, variant or fragment is acellular growth or proliferation promoting peptide, variant or fragment,respectively.

Homologs of Rice Protein 4 (SEQ ID NO: 6) include Oryza sativa JaponicaGroup, Brachipodium distachyon (SEQ ID NO: 540 to 542).

[SEQ ID NO: 7 (Pea Protein 3—P09918)]

Preferably, the peptide comprises a bioactive fragment of the protein ofSEQ ID NO: 7 or a homolog thereof, or a bioactive variant of thefragment.

Preferably, the peptide comprises a bioactive fragment of SEQ ID NO'S:283, or a bioactive variant of the fragment.

The invention also provides a composition comprising at least 1,preferably at least 2, preferably at least 3, preferably at least 4,preferably at least 5, preferably at least 6, preferably at least 7,preferably at least 8, preferably at least 9, or preferably at least 10bioactive peptides of the invention, each of which comprises a differentbioactive fragment of SEQ ID NO: 7 or a homolog thereof. Preferably, thecomposition comprises a bioactive fragment SEQ ID NO'S: 283, or abioactive variant of the fragment.

In one embodiment, the bioactive peptide, variant or fragment is acellular growth or proliferation promoting peptide, variant or fragment,respectively.

Homologs of Pea Protein 3 (SEQ ID NO: 7) include Medicago truncatula,Glycine soja, and Phaseolus vulgaris (SEQ ID NO: 543 to 545).

[SEQ ID NO: 8 (Pea Protein 4—P02857)]

Preferably, the peptide comprises a bioactive fragment of the protein ofSEQ ID NO: 8 or a homolog thereof, or a bioactive variant of thefragment.

Preferably, the peptide comprises a bioactive fragment selected from SEQID NO'S: 284-307, or a bioactive variant of the fragment.

The invention also provides a composition comprising at least 1,preferably at least 2, preferably at least 3, preferably at least 4,preferably at least 5, preferably at least 6, preferably at least 7,preferably at least 8, preferably at least 9, or preferably at least 10bioactive peptides of the invention, each of which comprises a differentbioactive fragment of SEQ ID NO: 8 or a homolog thereof. Preferably, thecomposition comprises a first bioactive peptide comprising a bioactivefragment selected from SEQ ID NO 284-307, and a second bioactive peptidecomprising a bioactive fragment selected from SEQ ID NO 284-307.

In one embodiment, the bioactive peptide, variant or fragment is acellular growth or proliferation promoting peptide, variant or fragment,respectively.

Homologs of Pea Protein 4 (SEQ ID NO: 8) include Vicia sativa, Vicianarbonesis and Cicer arietinum (SEQ ID NO: 546 to 548).

[SEQ ID NO: 9 (Pea Protein 5—P02855)]

Preferably, the peptide comprises a bioactive fragment of the protein ofSEQ ID NO: 9 or a homolog thereof, or a bioactive variant of thefragment.

Preferably, the peptide comprises a bioactive fragment selected from SEQID NO'S: 308-339, or a bioactive variant of the fragment.

The invention also provides a composition comprising at least 1,preferably at least 2, preferably at least 3, preferably at least 4,preferably at least 5, preferably at least 6, preferably at least 7,preferably at least 8, preferably at least 9, or preferably at least 10bioactive peptides of the invention, each of which comprises a differentbioactive fragment of SEQ ID NO: 9 or a homolog thereof. Preferably, thecomposition comprises a first bioactive peptide comprising a bioactivefragment selected from SEQ ID NO 308-339, and a second bioactive peptidecomprising a bioactive fragment selected from SEQ ID NO 308-339.

In one embodiment, the bioactive peptide, variant or fragment is acellular growth or proliferation promoting peptide, variant or fragment,respectively.

Homologs of Pea Protein 5 (SEQ ID NO: 9) include Lathyrus hirsutus,Lathyrus cicero, Lathyrus sativus (SEQ ID NO: 549 to 551).

[SEQ ID NO: 10 (Pea Protein 6—D3VNE1)]

Preferably, the peptide comprises a bioactive fragment of the protein ofSEQ ID NO: 10 or a homolog thereof, or a bioactive variant of thefragment.

Preferably, the peptide comprises the bioactive fragment SEQ ID NO: 340,or a bioactive variant of the fragment.

The invention also provides a composition comprising at least 1,preferably at least 2, preferably at least 3, preferably at least 4,preferably at least 5, preferably at least 6, preferably at least 7,preferably at least 8, preferably at least 9, or preferably at least 10bioactive peptides of the invention, each of which comprises a differentbioactive fragment of SEQ ID NO: 10 or a homolog thereof. Preferably,the composition comprises a first bioactive peptide comprising thebioactive fragment SEQ ID NO 340.

In one embodiment, the bioactive peptide, variant or fragment is acellular growth or proliferation promoting peptide, variant or fragment,respectively.

Homologs of Pea Protein 6 (SEQ ID NO: 10) include Medicago truncatula,Vicia peregrine, and Vicia lutea (SEQ ID NO: 552 to 554).

[SEQ ID NO: 11 (Rice Protein 5—P07728)]

Preferably, the peptide comprises a bioactive fragment of the protein ofSEQ ID NO: 11 or a homolog thereof, or a bioactive variant of thefragment.

Preferably, the peptide comprises a bioactive fragment selected from SEQID NO'S: 341-358, or a bioactive variant of the fragment.

The invention also provides a composition comprising at least 1,preferably at least 2, preferably at least 3, preferably at least 4,preferably at least 5, preferably at least 6, preferably at least 7,preferably at least 8, preferably at least 9, or preferably at least 10bioactive peptides of the invention, each of which comprises a differentbioactive fragment of SEQ ID NO: 11 or a homolog thereof. Preferably,the composition comprises a first bioactive peptide comprising abioactive fragment selected from SEQ ID NO 341-358, and a secondbioactive peptide comprising a bioactive fragment selected from SEQ IDNO 341-358.

In one embodiment, the bioactive peptide, variant or fragment is acellular growth or proliferation promoting peptide, variant or fragment,respectively.

Homologs of Rice Protein 5 (SEQ ID NO: 11) include Oryza sativa IndicaGroup, Zizania latifolia, Avena sativa (SEQ ID NO: 555 to 557).

[SEQ ID NO: 12 (Rice Protein 6—P07730)]

Preferably, the peptide comprises a bioactive fragment of the protein ofSEQ ID NO: 12 or a homolog thereof, or a bioactive variant of thefragment.

Preferably, the peptide comprises a bioactive fragment selected from SEQID NO'S: 359-401, or a bioactive variant of the fragment.

The invention also provides a composition comprising at least 1,preferably at least 2, preferably at least 3, preferably at least 4,preferably at least 5, preferably at least 6, preferably at least 7,preferably at least 8, preferably at least 9, or preferably at least 10bioactive peptides of the invention, each of which comprises a differentbioactive fragment of SEQ ID NO: 12 or a homolog thereof. Preferably,the composition comprises a first bioactive peptide comprising abioactive fragment selected from SEQ ID NO 359-401, and a secondbioactive peptide comprising a bioactive fragment selected from SEQ IDNO 359-401.

In one embodiment, the bioactive peptide, variant or fragment is acellular growth or proliferation promoting peptide, variant or fragment,respectively.

Homologs of Rice Protein 6 (SEQ ID NO: 12) include Oryza brachyantha,Brachipodium distachyon (SEQ ID NO: 558 to 560).

[SEQ ID NO: 13 (Rice Protein 7—Q0D7S0)]

Preferably, the peptide comprises a bioactive fragment of the protein ofSEQ ID NO: 13 or a homolog thereof, or a bioactive variant of thefragment.

Preferably, the peptide comprises a bioactive fragment selected from SEQID NO'S: 402-412, or a bioactive variant of the fragment.

The invention also provides a composition comprising at least 1,preferably at least 2, preferably at least 3, preferably at least 4,preferably at least 5, preferably at least 6, preferably at least 7,preferably at least 8, preferably at least 9, or preferably at least 10bioactive peptides of the invention, each of which comprises a differentbioactive fragment of SEQ ID NO: 13 or a homolog thereof. Preferably,the composition comprises a first bioactive peptide comprising abioactive fragment selected from SEQ ID NO 402-412, and a secondbioactive peptide comprising a bioactive fragment selected from SEQ IDNO 402-412.

In one embodiment, the bioactive peptide, variant or fragment is acellular growth or proliferation promoting peptide, variant or fragment,respectively.

Homologs of Rice Protein 7 (SEQ ID NO: 13) include Oryza sativa IndicaGroup, Zizania latifolia, Avena sativa (SEQ ID NO: 561 to 564).

[SEQ ID NO: 14 (Staphylococcus Aureus Protein 1—P0C1U8)]

Preferably, the peptide comprises a fragment of the protein of SEQ IDNO: 14 or a homolog thereof.

Preferably, the peptide comprises a bioactive fragment selected from SEQID NO'S: 413-417, or a bioactive variant of the fragment.

The invention also provides a composition comprising at least 1,preferably at least 2, preferably at least 3, preferably at least 4,preferably at least 5, preferably at least 6, preferably at least 7,preferably at least 8, preferably at least 9, or preferably at least 10bioactive peptides of the invention, each of which comprises a differentbioactive fragment of SEQ ID NO: 14 or a homolog thereof. Preferably,the composition comprises a first bioactive peptide comprising abioactive fragment selected from SEQ ID NO 413-417, and a secondbioactive peptide comprising a bioactive fragment selected from SEQ IDNO 413-417.

In one embodiment, the bioactive peptide, variant or fragment is acellular growth or proliferation promoting peptide, variant or fragment,respectively.

Homologs of Staphylococcus aureus Protein 1 (SEQ ID NO: 14) include:

>gi|580560623|gb|EVF84961.1| glutamyl endopeptidase [Staphylococcusaureus COAS6020]

>gi|580687002|gb|EVH10169.1| glutamyl endopeptidase [Staphylococcusaureus UCIM6080]

>gi|751815683|gb|KIN24957.1| glutamyl endopeptidase [Staphylococcusaureus MRSA_CVM43477]

>gi|781884797|dbj|BAR08486.1| glutamyl endopeptidase precursor[Staphylococcus aureus subsp. aureus]

>gi|781887762|dbj|BAR11210.1| glutamyl endopeptidase precursor[Staphylococcus aureus subsp. aureus]

The invention also provides a composition comprising at least one andpreferably a plurality of peptides of the invention, wherein each of thepeptides of the invention comprises a bioactive fragment of a proteindisclosed herein, for example selected from SEQ ID NO: 1 to 14 or ahomolog thereof, or a bioactive variant of the fragment.

Typically, the or each peptide of the invention comprises, a bioactivefragment selected from, SEQ ID NO: 15-505 and 546 to 704 and 717-775, ora bioactive variant of the fragment.

Typically, the or each peptide of the invention is selected from abioactive fragment selected from, SEQ ID NO: 15-505 and 546 to 704 and717-775, or a bioactive variant of the fragment.

The invention also provides a composition comprising at least one andpreferably a plurality of peptides of the invention, wherein the or eachof the peptides of the invention comprise a bioactive fragment of a peaor rice protein disclosed herein, typically selected from SEQ ID NO: 1-2and 7-10. Typically, the or each peptide of the invention is selectedfrom, or comprises a bioactive fragment selected from, SEQ ID NO: 15-215and 283 to 340, or a bioactive variant of the fragment.

The invention also provides a composition comprising at least one andpreferably a plurality of peptides of the invention, wherein the or eachof the peptides of the invention comprise a bioactive fragment of aprotein disclosed herein typically selected from SEQ ID NO: 3-6 and11-13. Typically, the or each peptide of the invention is selected from,or comprises a bioactive fragment selected from, SEQ ID NO: 216-282 and341-412, or a bioactive variant of the fragment.

Preferably, the composition comprises at least two distinct growthpromoting peptides of the invention.

Preferably, the composition comprises at least three distinct growthpromoting peptides of the invention.

Preferably, the composition comprises at least four distinct growthpromoting peptides of the invention.

Preferably, the composition comprises at least five distinct growthpromoting peptides of the invention.

Preferably, the composition comprises at least six distinct growthpromoting peptides of the invention.

Preferably, the composition comprises at least seven distinct growthpromoting peptides of the invention.

Preferably, the composition comprises at least eight distinct growthpromoting peptides of the invention.

Preferably, the composition comprises at least nine distinct growthpromoting peptides of the invention.

Preferably, the composition comprises at least ten distinct growthpromoting peptides of the invention.

In one embodiment, the composition comprises one or more of SEQ ID NO'S247 to 268.

In one embodiment, the composition comprises one or more of SEQ ID NO'S248, 249, 252, 253 and 257.

In one embodiment, the invention comprises a composition comprisingsubstantially all of fragments SEQ ID NO: 15-215 and 283-340, or growthpromoting variants of the fragments, or a mixture of the growthpromoting fragments and variants.

In one embodiment, the invention comprises a composition comprisingsubstantially all of fragments SEQ ID NO: 216-282 and 341-412, or growthpromoting variants of the fragments, or a mixture of the growthpromoting fragments and variants.

In one embodiment, the composition is enriched in peptides having amolecular weight of less than 10 KD.

In one embodiment, the composition is a powder.

The invention also relates to a plaster, bandage or dressing suitablefor application to a wound and comprising a peptide or composition ofthe invention.

The invention also relates to a man-made cell culture media comprising apeptide of the invention. The invention also relates to a man-made cellculture media comprising a composition of the invention. In oneembodiment, the cell culture media is formulated for culture ofeukaryotic cells. In one embodiment, the cell culture media isformulated for culture of prokaryotic cells.

The invention also relates to a plaster, bandage or dressing suitablefor application to a wound and comprising a peptide or composition ofthe invention.

The invention also relates to a comestible product comprising a growthpromoting peptide of the invention. Preferably the comestible product isman-made.

The invention also relates to a comestible product comprising acomposition of peptides of the invention. Preferably the comestibleproduct is man-made.

Preferably, the comestible product is a food product for human or animal(mammalian) or cellular consumption.

In one embodiment the man-made comestible product is a beverage, In oneembodiment the man-made comestible product is a bakery product. In oneembodiment the man-made comestible product is a dairy product. In oneembodiment the man-made comestible product is a snack product. In oneembodiment the man-made comestible product is a baked extruded foodproduct. In one embodiment the man-made comestible product is powderedmilk. In one embodiment the man-made comestible product is an infantformula product. In one embodiment the man-made comestible product is aconfectionery product. In one embodiment the man-made comestible productis a yoghurt. In one embodiment the man-made comestible product is ayoghurt drink. In one embodiment the man-made comestible product is anice cream product. In one embodiment the man-made comestible product isa frozen food product. In one embodiment the man-made comestible productis a breakfast cereal. In one embodiment the man-made comestible productis a bread. In one embodiment the man-made comestible product is aflavoured milk drink. In one embodiment the man-made comestible productis a confectionery bar. In one embodiment the man-made comestibleproduct is a tea or tea product. In one embodiment the man-madecomestible product is a based extruded snack product. In one embodimentthe man-made comestible product is a fried snack product. In oneembodiment the man-made comestible product is a nutritional supplement.In one embodiment the man-made comestible product is a sportsnutritional product. In one embodiment the man-made comestible productis a baby food product. In one embodiment the man-made comestibleproduct is a specialty food product for immunocompromised individuals.In one embodiment the man-made comestible product is a food forgeriatric patients.

The invention also relates to a peptide of the invention for use inpromoting growth of a cell.

The invention also relates to a peptide of the invention for use inpromoting growth of a cell culture.

The invention also relates to a peptide of the invention for use inpromoting growth of a tissue.

The invention also relates to a peptide of the invention for use inpromoting growth of dermal or epithelial tissue.

The invention also relates to a peptide of the invention for use inpromoting growth of skin.

The invention also relates to a peptide of the invention for use inpromoting growth of an organ.

The invention also relates to a peptide of the invention for use inpromoting growth of an organism.

The invention also relates to a composition of the invention for use inpromoting growth of a cell.

The invention also relates to a composition of the invention for use inpromoting growth of a cell culture.

The invention also relates to a composition of the invention for use inpromoting growth of a tissue.

The invention also relates to a composition of the invention for use inpromoting growth of epithelial tissue.

The invention also relates to a composition of the invention for use inpromoting growth of skin.

The invention also relates to a composition of the invention for use inpromoting growth of an organ.

The invention also relates to a composition of the invention for use inpromoting growth of an organism.

In one embodiment, the cell, tissue or organism has a normal pathology(for example ageing skin). In one embodiment of the invention, the cell,tissue or skin has abnormal pathology (for example tissue damaged due totrauma, drug use, or epithelial tissue in the GI tract damaged due to aninflammatory disorder).

The growth promoting uses may be in-vivo or in-vitro uses. The growthpromoting uses may involve administration to mammal externally (i.e. tothe skin) or internally (i.e. to the GI tract).

The invention also relates to a peptide of the invention for use inslowing or inhibiting ageing of human skin.

The invention also relates to a method of slowing or inhibiting ageingof human skin comprising a step of administering a peptide of theinvention to the human skin. Typically, the peptide of the invention isadministered topically to the skin. Administration may be by means of aplaster or patch or a formulation suitable for topical application.

The invention also relates to a composition of the invention for use inslowing or inhibiting ageing of human skin. The invention also relatesto a peptide of the invention for use in preventing or slowing ageing ofthe human skin.

The invention also relates to a method of slowing or inhibiting ageingof human skin comprising a step of administering a composition of theinvention to the human skin.

Typically, the composition of the invention is administered topically tothe skin.

The invention also relates to a peptide of the invention for use intreatment of a wound in a mammal.

The invention also relates to a composition of peptides of the inventionfor use in treatment of a wound in a mammal.

The invention also relates to a wound treatment composition or productof the invention for use in treatment of a wound in a mammal.

The invention also relates to a peptide of the invention for use intreatment or prevention of a disease or condition characterised bydamaged epithelial cells or tissue.

The invention also relates to a composition of peptides of the inventionfor use in treatment or prevention of a disease or conditioncharacterised by damaged dermal or epithelial cells or tissue.

In one embodiment, the disease or condition characterised by damageddermal or epithelial cells or tissue is selected from cancer, trauma Theinvention also relates to a peptide of the invention for use inmaintaining or restoring gut health in a mammal.

The invention also relates to a composition of peptides of the inventionfor use in maintaining or restoring gut health in in a mammal.

The invention also relates to a peptide of the invention for use inmaintaining or restoring muscle health (for example lean tissue mass) ina mammal.

The invention also relates to a composition of peptides of the inventionfor use in maintaining or restoring muscle health (for example leantissue mass) in in a mammal.

The invention also relates to a pharmaceutical composition comprising apeptide of the invention in combination with a pharmaceuticallyacceptable carrier.

The invention also relates to a pharmaceutical composition comprising acomposition of peptides of the invention in combination with apharmaceutically acceptable carrier.

The invention also relates to a peptide of the invention for use intreatment or prevention of an inflammatory disorder in a mammal.

The invention also relates to a composition of the invention for use intreatment or prevention of an inflammatory disorder in a mammal.

The invention also relates to a comestible product, for example a foodproduct comprising a peptide or composition of the invention, forexample a dairy or non-dairy product, a solid food or a beverage, a foodadditive or supplement. The dairy product may be a milk, a cheese, oryoghurt. In one embodiment, the food product is a snack bar. The foodproduct may comprise any amount of the composition of the invention, forexample from 0.1% to 30% (w/w).

The peptides of the invention are used in the topical cosmetic orpharmaceutical composition of this invention at cosmetically orpharmaceutically effective concentrations to achieve the desired effect;in a preferred form with regards to the total weight of the composition,between 0.00000001% (in weight) and 20% (in weight); preferably between0.000001% (in weight) and 15% (in weight), more preferably between0.0001% (in weight) and 10% (in weight) and even more preferably between0.0001% (in weight) and 5% (in weight). Ideally, the peptides of thepresent invention are preferably used from about 0.00001% w/w to about0.5% w/w [0.1 to 5000 ppm], and more preferably from 0.00005 w/w toabout 0.05 w/w [0.5 to 500 ppm], and most preferably from about 0.0001w/w to about 0.01 w/w of the composition [1 to 100 ppm]. Ideally, thepeptides of the present invention are preferably used from about 0.0001%w/w to about 0.004% w/w of the composition.

For compositions of peptides of the invention, a typical daily dosagemay be 0.2 g to 100 g. However, when administered as a food for specialmedicinal purpose, or medical food, the daily dosage may be 50-500 g perday.

The dosage of compositions of the invention for use in food products andfood supplements (i.e. comestible compositions) will be broadly in the0.2-100 g/day range. In one embodiment, the daily dosage is 1-10 g/day,ideally about 3-8 g/day. In one embodiment, the daily dosage is 10-20g/day. In one embodiment, the daily dosage is 20-30 g/day. In oneembodiment, the daily dosage is 30-40 g/day. In one embodiment, thedaily dosage is 10-100 g/day. In one embodiment, the daily dosage isabout 5 g/day, ideally about 3-8 g/day. In one embodiment, the dosage is2-1000 mg/day/kg body weight. In one embodiment, the dosage is 10-500mg/day/kg body weight. In one embodiment, the dosage is 10-100 mg/day/kgbody weight. In one embodiment, the dosage is 30-70 mg/day/kg bodyweight. The dosage of peptides of the invention for food supplements maybe 0.00001 mg-0.01 mg per day or dose.

The food product may be a Food for Specific Medicinal Purposes (FSMP)which is defined as foods that are specifically formulated, processedand intended for the dietary management of diseases, disorders ormedical conditions of individuals who are being treated under medicalsupervision. These foods are intended for the exclusive or partialfeeding of people whose nutritional requirements cannot be met by normalfoods.

The invention also relates to a man-made personal care compositioncomprising a peptide of the invention.

The invention also relates to a man-made personal care compositioncomprising a composition of peptides of the invention.

In one embodiment the personal care composition is a skincare product.In one embodiment the personal care composition is a product formulatedfor topical application to the skin of a human. In one embodiment thepersonal care composition is an anti-aging product. In one embodimentthe personal care composition is a dentrifice product. In one embodimentthe personal care composition is a perfumery product. In one embodimentthe personal care composition is a deodorant product. In one embodimentthe personal care composition is an anti-perspirant product. In oneembodiment the personal care composition is a soap. In one embodimentthe personal care composition is a liquid soap. In one embodiment thepersonal care composition is a cream. In one embodiment the personalcare composition is a lotion. In one embodiment the personal carecomposition is a gel. In one embodiment the personal care composition isa powder.

The invention also relates to a man-made wound treatment compositioncomprising a peptide of the invention. The invention also relates to aman-made wound treatment composition comprising a composition of theinvention. Typically, the wound treatment composition is formulated fortopical application to a wound. In one embodiment, the compositioncomprises a cream, gel, lotion, powder.

The invention also provides topical composition comprising a peptide ofthe invention. It will be appreciated that the topical composition maycomprise a plurality of peptides, fragments and/or variants. In oneembodiment, the topical composition comprises substantially all thepeptides. In one embodiment, the topical composition comprisessubstantially all the variants. The topical composition of the inventionmay be presented in a formulation selected from the group comprisingcreams, multiple emulsions, anhydrous compositions, aqueous dispersions,oils, milks, balsams, foams, lotions, gels, cream gels, hydro-alcoholicsolutions, hydro-glycolic solutions, cosmetic, personal care product,hydrogels, liniments, sera, soaps, dusting powder, paste, semi solidformulations, liniments, serums, shampoo, conditioner, ointments, anyrinse off formulation, talc, mousses, powders, sprays, aerosols,solutions, suspensions, emulsions, syrups, elixirs, polysaccharidefilms, patches, gel patches, bandages, an adhesive system, water-in-oilemulsions, oil-in-water emulsions, and silicone emulsions.

In an embodiment of the current invention, the emulsion contains a lipidor oil. The emulsion may be, but is not limited to, oil-in-water,water-in-oil, water-in-oil-in-water and oil-in-water-in-siliconeemulsions. The emulsion may contain a humectant. The emulsion maycontain an anti-foaming agent, such as silicone. The emulsion may haveany suitable viscosity. Emulsions may further contain an emulsifierand/or an anti-foaming agent. Methods of preparing an emulsion are knownto a person skilled in the art.

The topical composition of the invention may be incorporated into amedical device for administration. Such a device can include but is notlimited to a fabric, patch, bandage, gauge, sock, tight, underwear,dressing, glove, mask, adhesive patches, non-adhesive patches, occlusivepatches and microelectric patches or suitable adhesive system. In suchan embodiment, the device is in direct contact with the keratinous layersuch as the skin, thus releasing the peptides of the invention. It willbe understood that the topical composition may be incorporated in anysuitable form as detailed herein. For example, the topical compositionor peptides of the invention can be incorporated into the device or bepresent on the surface of the device or can be in a cream, gel or waxformulation or any suitable formulation defined herein and incorporatedinto the device or on the surface of the device. The device may beadapted for adhesion or attachment to the skin.

In one embodiment the device is adapted to release a constant quantityof the composition or the peptides of the invention. It will beunderstood that the amount of the composition contained in the sustainedrelease system will depend, for example, on where the composition is tobe administered, the kinetics and duration of the release of thecomposition of the invention, as well as the nature of the condition,disorder and/or disease to be treated and/or cared for. The device maybe such that the composition is released by biodegradation of thedevice, or by friction between the device and the body, due to bodilymoisture, the skin's pH or body temperature.

In an embodiment of the invention the topical composition may furthercomprise at least one cosmetically or pharmaceutically acceptableexcipient. Excipient may be used interchangeably with functionalingredient or additive. It will be understood that although the topicalcompositions of the current invention can be administered alone, theywill generally be administered in admixture with a cosmetic orpharmaceutical excipient. Cosmetically or pharmaceutically acceptableexcipient are well known in the art and any known excipient, may be usedprovided that it is suitable for topical administration and isdermatologically acceptable without undue toxicity, incompatibilityand/or allergic reaction.

Preferably any excipient included is present in trace amounts. Theamount of excipient included will depend on numerous factors, includingthe type of excipient used, the nature of the excipient, thecomponent(s) of the topical composition, the amount of active or peptidein the topical composition and/or the intended use of the topicalcomposition. The nature and amount of any excipient should notunacceptably alter the benefits of the peptides of this invention.

In an embodiment of the invention the excipient may be a suitablediluent, carrier, binder, lubricant, suspending agent, coating agent,preservative, stabilisers, dyes, vehicle, solubilising agent, base,emollient, emulsifying agent, fragrance, humectant, and/or surfactants.

Examples of suitable diluents include, but are not limited to, anydiluent disclosed in disclosed in US2014120131 or US2004132667. Examplesinclude ethanol, glycerol and water.

Examples of suitable carriers include, but are not limited to, lactose,starch, glucose, methyl cellulose, magnesium stearate, mannitol,sorbitol and any suitable carrier disclosed in US2014120131 orUS2004132667.

Examples of suitable binders include, but are not limited to, starch,gelatin, natural sugars such as glucose, anhydrous lactose, free-flowlactose, beta-lactose, corn sweeteners, natural and synthetic gums, suchas acacia, tragacanth or sodium alginate, carboxymethyl cellulose andpolyethylene glycol and any suitable binder disclosed in US2014120131 orUS2004132667.

Examples of suitable lubricants include, but are not limited to, sodiumoleate, sodium stearate, magnesium stearate, sodium benzoate, sodiumacetate, and sodium chloride and any suitable lubricant disclosed inUS2014120131 or US2004132667.

The carrier may be any suitable carried known in the art or disclosed inUS2014120131 or US2004132667. In some embodiments, the carrier mayinclude, but is not limited to, a liquid, such as water, oils orsurfactants, including those of petroleum, animal, plant or syntheticorigin, polymer, oil, such as peanut oil, mineral oil, castor oil,soybean oil, alcohol, polysorbates, sorbitan esters, ether sulfates,sulfates, betaines, glycosides, maltosides, fatty alcohols, nonoxynols,poloxamers, polyoxyethylenes, polyethylene glycols, dextrose, glycerol,or digitonin. It will be understood that the carrier will bedermatologically acceptable. Preferred carriers contain an emulsion suchas oil-in-water, water-in-oil, water-in-oil-in-water andoil-in-water-in-silicone emulsions. Emulsions may further contain anemulsifier and/or an anti-foaming agent.

In an embodiment of the invention, the topical composition may furthercomprise one or more additional ingredients. The topical composition ofthe invention may be administered consecutively, simultaneously orsequentially with the one or more other additional agents. Suchadditional ingredients may be those of benefit to include in a topicalcomposition, or of benefit depending on the intended use of the topicalcomposition. The additional ingredient may be active or functional orboth.

Examples of such additional ingredients include, but are not limited to,one or more of cleaning agents, conditioning agents, sunscreen, pigment,moisturiser, thickening agents, gelling agents, essential oil,astringents, pigments, anti-caking agent, anti-foaming agent, binders,additives, buffers, chelating agents, external analgesics, film formersor materials, bulking agents, polymers, opacifying agents, pH adjusters,propellants, reducing agents, sequestrants, skin bleaching andlightening agents, skin conditioning agents, aloe vera, healing agents,soothing agents, smoothing agents, pantothenic acid, treating agents,thickeners, vitamins. colourants, pharmaceuticals, antiseptic agents,antifoaming agents, buffering agents, astringents, polymers, pHadjuster, deodorant or any other dermatologically acceptable carrier orsurfactant.

It is to be understood that additional ingredients listed may providemore than one benefit. The classification given herein is for clarityand convenience only and not intended to limit the additional ingredientto that particular application or category listed.

Any additional ingredients should be suitable for application to theskin without undue toxicity, incompatibility and/or allergic reaction.

In some embodiments, the additional ingredient has glucose transportactivity or aids glucose transport activity. In some embodiments, theadditional ingredient has anti-inflammatory activity or aidsanti-inflammatory activity. In some embodiments, the additionalingredient has anti-aging activity or aids anti-aging activity. In someembodiments, the additional ingredient is for keratinous layer healthand/or development, skin health and/or development, and/or musclehealth, recovery and/or development. The active agent may be apharmacological enhancer. Such active agents are known and available onthe market. In such cases, the topical composition of the invention maybe administered consecutively, simultaneously or sequentially with theone or more other active agents.

In some embodiments, the additional ingredient may be farnesol ([2E,6E],-3, 7, 11,-trimethyl-2, 6, 10, dodecatrien-1-ol), phytantriol (3, 7,11, 15, tetramethylhexadecane-1, 2, 3,-triol), desquamation actives,enzymes, enzyme inhibitors, enzyme activators, botanical extracts andmarine extracts, anti-acne actives, anti-wrinkle or anti atrophyactives, anti-oxidant/radical scavengers, chelators, flavonoids,anti-inflammatory agents, anti-cellulite agents, topical anaesthetics,tanning actives, skin lightening agents, skin healing agents, bisabolol,antimicrobial or antifungal active, sunscreen actives, particulatematerial, conditioning agents, structuring agents, thickening agent,

The desquamation active may be any suitable agent that enhances the skinappearance or texture of the skin and is as disclosed in US2014120131 orUS2004132667.

Examples of anti-acne actives are as disclosed in US2014120131 orUS2004132667 and include, resorcinol, salicylic acid, erythromycin,zine, sulfur, benzoyl peroxides.

Examples of thickening agents are as disclosed in US2014120131 orUS2004132667 and include carboxylic acid polymers, crosslinkedpolyacrylate polymers, polyacrylamide polymers, polysaccharides.

Examples of conditioning agents are as disclosed in US2014120131 orUS2004132667 and include humectants, moisturiser or skin conditioner.

Examples of structuring agents are as disclosed in US2014120131 orUS2004132667 and include any agent that provide rheologicalcharacteristics to the composition and contributes to the stability ofthe composition.

Any suitable antimicrobial or antifungal active may be used and examplesare as disclosed in US2014120131 or US2004132667. Such actives arecapable of destroying microbes, preventing growth or action of microbes.Examples include but are not limited to β-lactam drugs, quinolone drugs,tetracycline, erythromycin, streptomycin sulfate, salicylic acid,benzoyl peroxide.

Examples of a particulate material include metallic oxide. Examples ofanti-cellulite agents include xanthine agents. Examples of tanningactives includes 1, 3-dihydroxy-2-propanone and those disclosed inUS2014120131 or US2004132667. Examples of topical anaesthetics includebenzocaine, lidocaine and bupivacaine and those disclosed inUS2014120131 or US2004132667.

Examples of skin lightening agents include any agent known in the artsuch as kojic acid, ascorbic acid and those disclosed in US2014120131 orUS2004132667.

Examples of sunscreen actives include any suitable organic or inorganicsunscreen active. Examples include metallic oxides,2-ethylhexyl-p-methoxycinnamate and those disclosed in US2014120131 orUS2004132667.

Examples of skin healing agents includes panthenoic acid as disclosed inUS2014120131 or US2004132667.

Examples of anti-inflammatory agents include any agent that enhances theskin appearance, tone or colour and include but are not limited tocorticosteroids, hydrocortisone, non-steroidal agents such as ibuprofenand aspirin and those disclosed in US2014120131 or US2004132667.

Examples of flavonoids includes flavanones, methoxy flavonones,unsubstituted chalcone and mixtures thereof and those disclosed inUS2014120131 or US2004132667.

Examples of enzymes include lipases, proteases, catalase, superoxide-dismutase, amylase, peroxidase, glucuronidase, ceramidases,hyaluronidases. Examples of enzyme inhibitors include trypsineinhibitors, Bowmann Birk inhibitors, chymotrypsin inhibitors, botanicalextracts, flavonoids, quercetin chalcone and those disclosed inUS2014120131 or US2004132667 and mixtures thereof. Examples of enzymeactivators include coenzyme A, Q10 (ubiquinone), glycyrrhizin,berberine, chrysin and those disclosed in US2014120131 or US2004132667and mixtures thereof

Examples of anti-wrinkle or anti atrophy actives include sulfurcontaining D and L amino acids, particular, N-acyl derivatives such asN-acetyl-L-cysteine, hydroxyl acids, phytic acid, lipoic acid,lysophosphatidic acid, skin peel agents, vitamin B₃, retinoids and thosedisclosed in US2014120131 or US2004132667 and mixtures thereof.

The anti-oxidant/radical scavenger agent may be any agent that is usefulfor providing protection against UV radiation or other environmentalagents which may cause skin damage such as those disclosed inUS2014120131 or US2004132667. Examples of anti-oxidant/radicalscavengers include ascorbic acid, its salts and derivatives (vitamin C),tocopherol its salts and derivatives (vitamin E), butylated hydroxylbenzoic acids and their salts, peroxides, gallic acids and alkyl esters,sorbic acid, lipoic acid, amines, lycine pidolate, arginine pilolate,nordihydroguaiaretic acid, bioflavonoids, curcumin, lysine, methionine,proline, superoxide dismutase, silymarin, tea extracts and mixturesthereof.

Examples of chelators include EDTA, NTA, hydoxamic acids, phytic acid,lactoferrin and those disclosed in US2014120131 or US2004132667 andmixtures thereof. A chelator means an agent capable of removing a metalion by forming a complex so that the metal ion cannot participate in orcatalyse chemical reactions. A chelator is useful for protection againstUV radiation or other environmental agents that can cause skin damage.

It will be appreciated that a plurality of additional ingredients may beadded. The amount of the additional ingredient may be from about 0.001%to about 50% weight of the composition, preferably, about 0.01% to about20%, preferably about 0.1% to about 10%, about 0.5% to about 10%, about1% to about 5%, preferably 2% weight of the composition. The amount ofadditional ingredient included will depend on numerous factors,including the type of additional ingredient used, the nature of theadditional ingredient, the component(s) of the topical composition, theamount of active or peptide in the topical composition and/or theintended use of the topical composition. The nature and amount of anyadditional ingredient should not unacceptably alter the benefits of thepeptides of this invention.

The topical composition may be alcohol free.

In some embodiments of the invention, the composition further comprisesone or more additional active agents, in addition to the peptide of theinvention (also known as the active of the composition). In addition, oralternatively, the composition may be administered with one or moreother additional active agents. Typical said additional active agent ispresent in trace amounts only. In some embodiments, there may be noadditional active agent present in the composition. The amount ofadditional active agent included will depend on numerous factors,including the type of additional active agent used, the nature of theadditional active agent, the component(s) of the topical composition,the amount of active or peptide in the topical composition and/or theintended use of the topical composition. The nature and amount of anyadditional active agent should not unacceptably alter the benefits ofthe peptides of this invention.

It is to be understood that an ingredient that is considered to be an“active” ingredient in one product may be a “functional” or “excipient”ingredient in another and vice versa. It will also be appreciated thatsome ingredients play a dual role as both an active ingredient and as afunctional or excipient ingredient.

Examples of the additional active agents include glucose transportpromoting drugs, skin supplement, agent for treatment and/or care of theskin, anti-inflammatory agent, an anti-aging agent, a cellular growthpromoting agent and pharmacological enhancers. Such agents are wellknown in the art and it will be appreciated that any suitable additionalactive agent may be used. Additional active agents for treatment and/orcare of the skin may include collagen synthesis agents, retinoids,exfoliating agents, anti-cellulite agents, elastase inhibiting agents,melanin synthesis stimulating or inhibiting agents, self-tanning agents,antiaging agents, antimicrobial agents, antifungal agents, fungistaticagents, bactericidal agents, and healing agents. Active agents alsoinclude anti-inflammatory agents.

Any additional active agent should be suitable for application to theskin without undue toxicity, incompatibility and/or allergic reaction.

It will be understood that the classification given herein is forclarity and convenience only and not intended to limit the additionalingredient, excipient, or active to that particular application orcategory listed.

In a particularly preferred embodiment, the methods and uses of theinvention involve administration of a peptide or composition of theinvention in combination with one or more other active agents, forexample, existing growth promoting drugs or pharmacological enhancersavailable on the market. In such cases, the compounds of the inventionmay be administered consecutively, simultaneously or sequentially withthe one or more other active agents.

The effect of the current invention is accomplished by topicalapplication or administration of the topical composition of theinvention described herein to a person, animal or a patient in need oftreatment or care. Topical delivery preferably means delivery to akeratinous layer such as the skin, hair and/or nails, but can also meandelivery to a body lumen lined with epithelial cells, for example thelungs or airways, the gastrointestinal tract, the buccal cavity. Theeffect may be confined to the surface of the skin or may be within theskin or a combination of both.

The topical composition of the invention is administered in acosmetically or pharmaceutically effective amount. In other words, in anamount that is non-toxic but sufficient amount to provide the desiredeffect. It will be appreciated that a person skilled in the art would becapable of determining an appropriate dose of the topical compositionsof the invention to administer without undue experimentation.Alternatively, a physician will determine the actual dose that is mostsuitable for a patient depending on the particular condition, disease ordisorder to be treated or cared for and the age, body weight and/orhealth of the person. It will depend on a variety of factors includingthe activity of the specific compound employed, the metabolic stabilityand length of action of that compound, the age, body weight, generalhealth, sex, diet, mode and time of administration, rate of excretion,drug combination, the severity of the particular condition, and theindividual undergoing therapy. There can, of course, be individualinstances where higher or lower dosage ranges are merited, and such arewithin the scope of this invention. For example, the composition may beadministered at a dose of from 0.01 to 50 mg/kg body weight, such asfrom 0.1 to 30 mg/kg, more preferably from 0.1 to 20 mg/kg body weight,more preferably from 0.1 to 10 mg/kg body weight, preferably 0.1 to 5mg/kg body weight. In an exemplary embodiment, one or more doses of 10to 300 mg/day or more preferably, 10 to 150 mg/day, will be administeredto the patient. The amount and the frequency is as best suited to thepurpose. The frequency of application or administration can varygreatly, depending on the needs of each subject, with a recommendationof an application or administration range from once a month to ten timesa day, preferably from once a week to four times a day, more preferablyfrom three times a week to three times a day, even more preferably onceor twice a day.

In preferred embodiments, repeated use of the topical composition isprovided.

The topical composition may be applied by, but not limited to, rubbing,or massaging into the keratinous tissue, skin or area of the body to betreated or cared for. In some embodiments, the composition is left on ornot removed from the area of the body. In other embodiments, thecomposition is removed after a period of time, such as, but not limitedto, from about 2 minutes to 60 minutes, from about 5 minutes to about 30minutes, preferably from about 10 minutes to about 20 minutes. Thecomposition may be removed immediately after application. In someembodiments of the current invention, the composition of the inventionmay be applied to an area to be treated by means to achieve a greaterpenetration of the composition and/or peptide of the invention, such as,but not limited to, iontophoresis, sonophoresis, electroporation,microelectric patches, mechanical pressure, osmotic pressure gradient,occlusive cure, microinjections or needle-free injections by means ofpressure, such as injections by oxygen pressure, or any combinationthereof.

The peptides of the invention are used in the topical cosmetic orpharmaceutical composition of this invention at cosmetically orpharmaceutically effective concentrations to achieve the desired effect;in a preferred form with regards to the total weight of the composition,between 0.00000001% (in weight) and 20% (in weight); preferably between0.000001% (in weight) and 15% (in weight), more preferably between0.0001% (in weight) and 10% (in weight) and even more preferably between0.0001% (in weight) and 5% (in weight).

In some embodiments of the current invention, the composition may bedelivered via any one of liposomes, mixed liposomes, oleosomes,niosomes, ethosomes, millicapsules, capsules, macrocapsules,nanocapsules, nanostructured lipid carriers, sponges, cyclodextrins,vesicles, micelles, mixed micelles of surfactants,surfactant-phospholipid mixed micelles, millispheres, spheres,lipospheres, particles, nanospheres, nanoparticles, milliparticles,solid nanopartciles as well as microemulsions including water-in-oilmicroemulsions with an internal structure of reverse micelle andnanoemulsions microspheres, microparticles.

A variety of methods are available for preparing liposomes. See, e.g.,Szoka et al., Ann. Rev. Biophys. Bioeng. 9:467 (1980), U.S. Pat. Nos.4,186,183, 4,217,344, 4,235,871, 4,261,975, 4,485,054, 4,501,728,4,774,085, 4,837,028, 4,235,871, 4,261,975, 4,485,054, 4,501,728,4,774,085, 4,837,028, 4,946,787, PCT Publication No. WO 91/17424, Deamer& Bangham, Biochim. Biophys. Acta 443:629-634 (1976); Fraley, et al.,PNAS 76:3348-3352 (1979); Hope et al., Biochim. Biophys. Acta 812:55-65(1985); Mayer et al., Biochim. Biophys. Acta 858:161-168 (1986);Williams et al., PNAS 85:242-246 (1988); Liposomes (Ostro (ed.), 1983,Chapter 1); Hope et al., Chem. Phys. Lip. 40:89 (1986); Gregoriadis,Liposome Technology (1984) and Lasic, Liposomes: from Physics toApplications (1993)). Suitable methods include, for example, sonication,extrusion, high pressure/homogenization, microfluidization, detergentdialysis, calcium-induced fusion of small liposome vehicles and etherfusion methods, all of which are well known in the art.

These delivery systems may be adapted to achieve a greater penetrationof the compound and/or peptides of the invention. This may improvepharmacokinetic and pharmacodynamics properties. The delivery system maybe a sustained release system wherein the compound or peptide of theinvention is gradually released during a period of time and preferablywith a constant release rate over a period of time. The delivery systemsare prepared by methods known in the art. The amount of peptidecontained in the sustained release system will depend on where thecomposition is to be delivered and the duration of the release as wellas the type of the condition, disease and/or disorder to be treated orcared for.

The topical composition of the invention may be for human or animalusage in human and veterinary medicine.

The topical composition of the invention may be used for pharmaceutical,personal care and/or cosmetic uses.

The composition can be used to treat or care for any disease, disorderor condition of the skin, including but not limited to, psoriasis,dermatitis, allergic dermatitis, eczema, spongiosis, edema, skin cancer,ulcers, acne, scars, cellulitis, elastosis, keratosis, rosacea, varicoseveins, inflammatory disorders.

The topical composition may be used to for treating or caring forvisible signs of aging including but not limited to wrinkles, stretchmarks and dark circles, dryness, fine lines, age spots, red blotches,sagging skin, and conditions caused by sun exposure including sunburn,stress, pollution and/diet. The topical composition may also be used fordelaying, slowing or inhibiting the skins or the onset of aging. Thecomposition may be administered by a medical device, such as a plasteror a patch as described herein.

The topical composition may be used to treat or care for a wound in amammal. In another embodiment, the topical composition is for use in thetreatment or prevention of a disease or condition characterised bydamaged epithelial cells or tissue, and/or damaged dermal or epithelialcells or tissue. The disease may be but is not limited to cancer andtrauma.

The topical composition may be used to treat or care for any musclecondition, to improve, muscle status in a mammal, to promote recovery ofmuscle, typically following exercise, to maintain or restore musclehealth (for example lean tissue mass) in a mammal, to enhance physicalperformance, in treatment or prevention of a disease or conditioncharacterised by lethargy or low energy levels.

The topical composition may be used to promote growth of a tissue,promote growth of epithelial tissue, promote growth of skin, promotegrowth of an organ, promote growth of an organism. The skin can have anormal pathology and/or an abnormal pathology.

The topical composition may also be used to treat or care for anyinflammatory disorder.

A further aspect of the invention relates to a pharmaceuticalcomposition comprising a peptide of the invention or a composition ofpeptides of the invention, admixed with one or more pharmaceuticallyacceptable diluents, excipients or carriers. Even though the peptidesand compositions of the present invention can be administered alone,they will generally be administered in admixture with a pharmaceuticalcarrier, excipient or diluent, particularly for human therapy. Thepharmaceutical compositions may be for human or animal usage in humanand veterinary medicine. Examples of such suitable excipients for thevarious different forms of pharmaceutical compositions described hereinmay be found in the “Handbook of Pharmaceutical Excipients, 2^(nd)Edition, (1994), Edited by A Wade and PJ Weller. In particular,formulations for topical delivery are described in Topical drug deliveryformulations edited by David Osborne and Antonio Aman, Taylor & Francis,the complete contents of which are incorporated herein by reference.Acceptable carriers or diluents for therapeutic use are well known inthe pharmaceutical art, and are described, for example, in Remington'sPharmaceutical Sciences, Mack Publishing Co. (A. R. Gennaro edit. 1985).Examples of suitable carriers include lactose, starch, glucose, methylcellulose, magnesium stearate, mannitol, sorbitol and the like. Examplesof suitable diluents include ethanol, glycerol and water. The choice ofpharmaceutical carrier, excipient or diluent can be selected with regardto the intended route of administration and standard pharmaceuticalpractice. The pharmaceutical compositions may comprise as, or inaddition to, the carrier, excipient or diluent any suitable binder(s),lubricant(s), suspending agent(s), coating agent(s), solubilisingagent(s). Examples of suitable binders include starch, gelatin, naturalsugars such as glucose, anhydrous lactose, free-flow lactose,beta-lactose, corn sweeteners, natural and synthetic gums, such asacacia, tragacanth or sodium alginate, carboxymethyl cellulose andpolyethylene glycol. Examples of suitable lubricants include sodiumoleate, sodium stearate, magnesium stearate, sodium benzoate, sodiumacetate, sodium chloride and the like.

Preservatives, stabilizers, dyes and even flavouring agents may beprovided in the pharmaceutical composition. Examples of preservativesinclude sodium benzoate, sorbic acid and esters of p-hydroxybenzoicacid. Antioxidants and suspending agents may be also used.

The peptide or composition of the invention may be adapted for topical,oral, rectal, parenteral, intramuscular, intraperitoneal,intra-arterial, intrabronchial, subcutaneous, intradermal, intravenous,nasal, vaginal, buccal or sublingual routes of administration. For oraladministration, particular use is made of compressed tablets, pills,tablets, gellules, drops, and capsules. Preferably, these compositionscontain from 1 to 250 mg and more preferably from 10-100 mg, of activeingredient per dose. Other forms of administration comprise solutions oremulsions which may be injected intravenously, intra-arterial,subcutaneously, intradermally, intraperitoneally or intramuscularly, andwhich are prepared from sterile or sterilisable solutions. Thepharmaceutical compositions of the present invention may also be in formof suppositories, vaginal rings, pessaries, suspensions, emulsions,lotions, ointments, creams, gels, sprays, solutions or dusting powders.The composition of the invention may be formulated for topical delivery.Topical delivery generally means delivery to the skin, but can also meandelivery to a body lumen lined with epithelial cells, for example thelungs or airways, the gastrointestinal tract, the buccal cavity. Inparticular, formulations for topical delivery are described in Topicaldrug delivery formulations edited by David Osborne and Antonio Aman,Taylor & Francis, the complete contents of which are incorporated hereinby reference. Compositions or formulations for delivery to the airwaysare described in O'Riordan et al (Respir Care, 2002, November 47),EP2050437, WO2005023290, US2010098660, and US20070053845. Compositionand formulations for delivering active agents to the iluem, especiallythe proximal iluem, include microparticles and microencapsulates wherethe active agent is encapsulated within a protecting matrix formed ofpolymer or dairy protein that is acid resistant but prone to dissolutionin the more alkaline environment of the ileum. Examples of such deliverysystems are described in EP1072600.2 and EP13171757.1. An alternativemeans of transdermal administration is by use of a skin patch. Forexample, the active ingredient can be incorporated into a creamconsisting of an aqueous emulsion of polyethylene glycols or liquidparaffin. The active ingredient can also be incorporated, at aconcentration of between 1 and 10% by weight, into an ointmentconsisting of a white wax or white soft paraffin base together with suchstabilisers and preservatives as may be required.

Injectable forms may contain between 10-1000 mg, preferably between10-250 mg, of active ingredient per dose.

Compositions may be formulated in unit dosage form, i.e., in the form ofdiscrete portions containing a unit dose, or a multiple or sub-unit of aunit dose.

A person of ordinary skill in the art can easily determine anappropriate dose of one of the instant compositions to administer to asubject without undue experimentation. Typically, a physician willdetermine the actual dosage which will be most suitable for anindividual patient and it will depend on a variety of factors includingthe activity of the specific compound employed, the metabolic stabilityand length of action of that compound, the age, body weight, generalhealth, sex, diet, mode and time of administration, rate of excretion,drug combination, the severity of the particular condition, and theindividual undergoing therapy. The dosages disclosed herein areexemplary of the average case. There can of course be individualinstances where higher or lower dosage ranges are merited, and such arewithin the scope of this invention. Depending upon the need, the agentmay be administered at a dose of from 0.01 to 30 mg/kg body weight, suchas from 0.1 to 10 mg/kg, more preferably from 0.1 to 1 mg/kg bodyweight. In an exemplary embodiment, one or more doses of 10 to 300mg/day or more preferably, 10 to 150 mg/day, will be administered to thepatient for the treatment of an inflammatory disorder.

In a particularly preferred embodiment, the methods and uses of theinvention involve administration of a peptide or composition of theinvention in combination with one or more other active agents, forexample, existing anti-inflammatory drugs or pharmacological enhancersavailable on the market. In such cases, the compounds of the inventionmay be administered consecutively, simultaneously or sequentially withthe one or more other active agents.

In one embodiment of the invention, the peptide of the invention may beadministered in the form of a conjugate comprising the peptide, and mayoptionally include a linker, and a partner molecule, for example aprotein such as an antibody molecule intended to increase the half-lifeof the conjugate in-vivo. In one embodiment, the peptide may be modifiedto substitute one or more amino acids with amino acids employed toattach partner molecules.

For example, an amino acid may be substituted with a lysine residue forthe purpose of conjugating a partner molecule such as a PEG molecule.

Definitions

All publications, patents, patent applications and other referencesmentioned herein are hereby incorporated by reference in theirentireties for all purposes as if each individual publication, patent orpatent application were specifically and individually indicated to beincorporated by reference and the content thereof recited in full.

Where used herein and unless specifically indicated otherwise, thefollowing terms are intended to have the following meanings in additionto any broader (or narrower) meanings the terms might enjoy in the art:

Unless otherwise required by context, the use herein of the singular isto be read to include the plural and vice versa. The term “a” or “an”used in relation to an entity is to be read to refer to one or more ofthat entity. As such, the terms “a” (or “an”), “one or more,” and “atleast one” are used interchangeably herein.

As used herein, the term “comprise,” or variations thereof such as“comprises” or “comprising,” are to be read to indicate the inclusion ofany recited integer (e.g. a feature, element, characteristic, property,method/process step or limitation) or group of integers (e.g. features,element, characteristics, properties, method/process steps orlimitations) but not the exclusion of any other integer or group ofintegers. Thus, as used herein the term “comprising” is inclusive oropen-ended and does not exclude additional, unrecited integers ormethod/process steps.

As used herein, the term “disease” is used to define any abnormalcondition that impairs physiological function and is associated withspecific symptoms. The term is used broadly to encompass any disorder,illness, abnormality, pathology, sickness, condition or syndrome inwhich physiological function is impaired irrespective of the nature ofthe aetiology (or indeed whether the aetiological basis for the diseaseis established). It therefore encompasses conditions arising frominfection, trauma, injury, surgery, radiological ablation, poisoning ornutritional deficiencies.

As used herein, the term “treatment” or “treating” refers to anintervention (e.g. the administration of an agent to a subject) whichcures, ameliorates or lessens the symptoms of a disease or removes (orlessens the impact of) its cause(s) (for example, the reduction inaccumulation of pathological levels of lysosomal enzymes). In this case,the term is used synonymously with the term “therapy”.

Additionally, the terms “treatment” or “treating” refers to anintervention (e.g. the administration of an agent to a subject) whichprevents or delays the onset or progression of a disease or reduces (oreradicates) its incidence within a treated population. In this case, theterm treatment is used synonymously with the term “prophylaxis”.

As used herein, an effective amount or a therapeutically effectiveamount of an agent defines an amount that can be administered to asubject without excessive toxicity, irritation, allergic response, orother problem or complication, commensurate with a reasonablebenefit/risk ratio, but one that is sufficient to provide the desiredeffect, e.g. the treatment or prophylaxis manifested by a permanent ortemporary improvement in the subject's condition. The amount will varyfrom subject to subject, depending on the age and general condition ofthe individual, mode of administration and other factors. Thus, while itis not possible to specify an exact effective amount, those skilled inthe art will be able to determine an appropriate “effective” amount inany individual case using routine experimentation and background generalknowledge. A therapeutic result in this context includes eradication orlessening of symptoms, reduced pain or discomfort, prolonged survival,improved mobility and other markers of clinical improvement. Atherapeutic result need not be a complete cure.

The term “human or animal” should be understood to means humans ormammalian or non-mammalian animals such as fish.

The term “composition” should be understood to mean a composition ofmatter made by the hand of man and not occurring in nature. Exemplarycompositions include food compositions, beverage compositions,pharmaceutical compositions, nutritional supplement compositions,personal care compositions and healthcare compositions.

The term “peptide” used herein refers to a polymer composed of 5 to 50amino acid monomers typically via peptide bond linkage. Peptides(including fragments and variants thereof) of and for use in theinvention may be generated wholly or partly by chemical synthesis or byexpression from nucleic acid. For example, the peptides of and for usein the present invention can be readily prepared according towell-established, standard liquid or, preferably, solid-phase peptidesynthesis methods known in the art (see, for example, J. M. Stewart andJ. D. Young, Solid Phase Peptide Synthesis, 2nd edition, Pierce ChemicalCompany, Rockford, Ill. (1984), in M. Bodanzsky and A. Bodanzsky, ThePractice of Peptide Synthesis, Springer Verlag, New York (1984). Whennecessary, any of the peptides employed in the invention can bechemically modified to increase their stability. A chemically modifiedpeptide or a peptide analog includes any functional chemical equivalentof the peptide characterized by its increased stability and/or efficacyin vivo or in vitro in respect of the practice of the invention. Theterm peptide analog also refers to any amino acid derivative of apeptide as described herein. A peptide analog can be produced byprocedures that include, but are not limited to, modifications to sidechains, incorporation of unnatural amino acids and/or their derivativesduring peptide synthesis and the use of cross-linkers and other methodsthat impose conformational constraint on the peptides or their analogs.Examples of side chain modifications include modification of aminogroups, such as by reductive alkylation by reaction with an aldehydefollowed by reduction with NaBH₄; amidation with methylacetimidate;acetylation with acetic anhydride; carbamylation of amino groups withcyanate; trinitrobenzylation of amino groups with 2, 4, 6,trinitrobenzene sulfonic acid (TNBS); alkylation of amino groups withsuccinic anhydride and tetrahydrophthalic anhydride; and pyridoxylationof lysine with pyridoxa-5′-phosphate followed by reduction with NABH₄.The guanidino group of arginine residues may be modified by theformation of heterocyclic condensation products with reagents such as2,3-butanedione, phenylglyoxal and glyoxal. The carboxyl group may bemodified by carbodiimide activation via o-acylisourea formation followedby subsequent derivatization, for example, to a corresponding amide.Sulfhydryl groups may be modified by methods, such as carboxymethylationwith iodoacetic acid or iodoacetamide; performic acid oxidation tocysteic acid; formation of mixed disulphides with other thiol compounds;reaction with maleimide; maleic anhydride or other substitutedmaleimide; formation of mercurial derivatives using4-chloromercuribenzoate, 4-chloromercuriphenylsulfonic acid,phenylmercury chloride, 2-chloromercuric-4-nitrophenol and othermercurials; carbamylation with cyanate at alkaline pH. Tryptophanresidues may be modified by, for example, oxidation withN-bromosuccinimide or alkylation of the indole ring with2-hydroxy-5-nitrobenzyl bromide or sulphonyl halides. Tryosine residuesmay be altered by nitration with tetranitromethane to form a3-nitrotyrosine derivative. Modification of the imidazole ring of ahistidine residue may be accomplished by alkylation with iodoacetic acidderivatives or N-carbethoxylation with diethylpyrocarbonate. Examples ofincorporating unnatural amino acids and derivatives during peptidesynthesis include, but are not limited to, use of norleucine, 4-aminobutyric acid, 4-amino-3-hydroxy-5-phenylpentanoic acid, 6-aminohexanoicacid, t-butylglycine, norvaline, phenylglycine, ornithine, sarcosine,4-amino-3-hydroxy-6-methylheptanoic acid, 2-thienyl alanine and/orD-isomers of amino acids. Peptide structure modification includes thegeneration of retro-inverso peptides comprising the reversed sequenceencoded by D-amino acids.

The term “modified peptide” is used interchangeably with the termderivative of the peptide. The modified peptide includes a peptide whichhas been substituted with one or more groups as defined herein. Themodification may be any modified that provides the peptides and or thecomposition of the invention with an increased ability to penetrate acell. The modification may be any modification that increases thehalf-life of the composition or peptides of the invention. In oneembodiment, the group is a protecting group. The protecting group may bean N-terminal protecting group, a C-terminal protecting group or aside-chain protecting group. The peptide may have one or more of theseprotecting groups. The person skilled in the art is aware of suitabletechniques to react amino acids with these protecting groups. Thesegroups can be added by preparation methods known in the art, for examplethe methods as outlined in paragraphs [0104] to [0107] of US2014120141.The groups may remain on the peptide or may be removed. The protectinggroup may be added during synthesis. In an embodiment of the inventionthe peptides may be substituted with a group selected from one or morestraight chain or branched chain, long or short chain, saturated, orunsaturated, substituted with a hydroxyl, amino, amino acyl, sulfate orsulphide group or unsubstituted having from 1 to 29 carbon atoms. N-acylderivatives include acyl groups derived from acetic acid, capric acid,lauric acid, myristic acid, octanoic acid, palmitic acid, stearic acid,behenic acid, linoleic acid, linolenic acid, lipoic acid, oleic acid,isosteric acid, elaidoic acid, 2-ethylhexaneic acid, coconut oil fattyacid, tallow fatty acid, hardened tallow fatty acid, palm kernel fattyacid, lanolin fatty acid or similar acids. These may be substituted orunsubstituted. When substituted they are preferably substituted withhydroxyl, or sulphur containing groups such as but not limited to SO₃H,SH, or S—S. In an embodiment of the current invention, the peptide isR₁—X—R₂. R₁ and/or R₂ groups respectively bound to the amino-terminal(N-terminal) and carboxyl-terminal (C-terminal) of the peptide sequence.In one embodiment, the peptide is R₁—X. Alternatively, the peptide isX—R₂. Preferably, R₁ is H, C₁₋₄ alkyl, acetyl, benzoyl ortrifluoroacetyl; X is the peptide of the invention; R₂ is OH or NH₂. Inan embodiment, R₁ is selected from the group formed by H, a non-cyclicsubstituted or unsubstituted aliphatic group, substituted orunsubstituted alicyclyl, substituted or unsubstituted heterocyclyl,substituted or unsubstituted heteroarylalkyl, substituted orunsubstituted aryl, substituted or unsubstituted aralkyl,Tert-butyloxycarbonyl, 9-fluorenylmethyloxycarbonyl (Fmoc) and R₅—CO—,wherein R₅ is selected from the group formed by H, a non-cyclicsubstituted or unsubstituted aliphatic group, substituted orunsubstituted alicyclyl, substituted or unsubstituted aryl, substitutedor unsubstituted aralkyl, substituted or unsubstituted heterocyclyl andsubstituted or unsubstituted heteroarylalkyl; R₂ is selected from thegroup formed by —NR₃R₄, —OR₃ and —SR₃, wherein R₃ and R₄ areindependently selected from the group formed by H, a non-cyclicsubstituted or unsubstituted aliphatic group, substituted orunsubstituted alicyclyl, substituted or unsubstituted heterocyclyl,substituted or unsubstituted heteroarylalkyl, substituted orunsubstituted aryl, and substituted or unsubstituted aralkyl; and withthe condition that R₁ and R₂ are not α-amino acids. In accordance withanother preferred embodiment, R₂ is —NR₃R₄, —OR₃ or —SR₃ wherein R₃ andR₄ are independently selected from the group formed by H, substituted orunsubstituted C₁-C₂₄ alkyl, substituted or unsubstituted C₂-C₂₄ alkenyl,Tert-butyloxycarbonyl, 9-fluorenylmethyloxycarbonyl (Fmoc), substitutedor unsubstituted C₂-C₂₄ alkynyl, substituted or unsubstituted C₃-C₂₄cycloalkyl, substituted or unsubstituted C₅-C₂₄ cycloalkenyl,substituted or unsubstituted C₈-C₂₄ cycloalkynyl, substituted orunsubstituted C₆-C₃₀ aryl, substituted or unsubstituted C₇-C₂₄ aralkyl,substituted or unsubstituted heterocyclyl ring of 3-10 members, andsubstituted or unsubstituted heteroarylalkyl of 2 to 24 carbon atoms and1 to 3 atoms other than carbon wherein the alkyl chain is of 1 to 6carbon atoms. Optionally, R₃ and R₄ can be bound by a saturated orunsaturated carbon-carbon bond, forming a cycle with the nitrogen atom.More preferably R₂ is —NR₃R₄ or —OR₃, wherein R₃ and R₄ areindependently selected from the group formed by H, substituted orunsubstituted C₁-C₂₄ alkyl, substituted or unsubstituted C₂-C₂₄ alkenyl,substituted or unsubstituted C₂-C₂₄ alkynyl, substituted orunsubstituted C₃-C₁₀ cycloalkyl, substituted or unsubstituted C₆-C₁₅aryl and substituted or unsubstituted heterocyclyl of 3-10 members,substituted or unsubstituted heteroarylalkyl with a ring of 3 to 10members and an alkyl chain of 1 to 6 carbon atoms. More preferably R₃and R₄ are selected from the group formed by H, methyl, ethyl, hexyl,dodecyl, or hexadecyl. Even more preferably R₃ is H and R₄ is selectedfrom the group formed by H, methyl, ethyl, hexyl, dodecyl, or hexadecyl.In accordance with an even more preferred embodiment, R₂ is selectedfrom —OH and —NH₂.

In accordance with another embodiment of this invention R₁ is selectedfrom the group formed by H, acetyl, lauroyl, myristoyl or palmitoyl, andR₂ is —NR₃R₄ or —OR₃ wherein R₃ and R₄ are independently selected fromH, methyl, ethyl, hexyl, dodecyl and hexadecyl, preferably R₂ is —OH or—NH₂. More preferably, R₁ is acetyl or palmitoyl and R₂ is —NH₂. In apreferred embodiment, the acyl group is bound to the N-terminal end ofat least one amino acid of the peptide. In an embodiment of theinvention, the peptide is modified to comprise a side chain protectinggroup. The side chain protecting group may be one or more of the groupcomprising benzyl or benzyl based groups, t-butyl-based groups,benzyloxy-carbonyl (Z) group, and allyloxycarbonyl (alloc) protectinggroup. The side chain protecting group may be derived from an achiralamino acid such as achiral glycine. The use of an achiral amino acidhelps to stabilise the resultant peptide and also facilitate the facilesynthesis route of the present invention. Preferably, the peptidefurther comprises a modified C-terminus, preferably an amidatedC-terminus. The achiral residue may be alpha-aminoisobutyric acid(methylalaine). It will be appreciated that the specific side chainprotecting groups used will depend on the sequence of the peptide andthe type of N-terminal protecting group used.

“Conjugate”: In one embodiment of the invention the peptide isconjugated, linked or fused to a binding partner, for example one ormore polyethylene glycol polymers or other compounds, such as molecularweight increasing compounds or lipophilic groups. The molecular weightincreasing compound is any compound that will increase the molecularweight, typically by 10% to 90%, or 20% to 50% of the resultingconjugate and may have a molecular weight of between 200 and 20,000,preferably between 500 and 10,000. The molecular weight increasingcompound may be PEG, any water-soluble (amphiphilic or hydrophilic)polymer moiety, homo or co-polymers of PEG, a monomethyl-substitutedpolymer of PEG (mPEG) and polyoxyethylene glycerol (POG), polyaminoacids such as poly-lysine, poly-glutamic acid, poly-aspartic acid,particular those of L conformation, pharmacologically inactive proteinssuch as albumin, gelatin, a fatty acid, polysaccharide, a lipid aminoacid and dextran. The polymer moiety may be straight chained or branchedand it may have a molecular weight of 500 to 40000 Da, 5000 to 10000 Da,10000 to 5000, Da. The compound (binding partner) may be any suitablecell penetrating compound, such as tat peptide, penetratin, pep-1. Thecompound (binding partner) may be an antibody molecule. The compound(binding partner) may be a lipophilic moiety or a polymeric moiety. Thelipophilic substituent and polymeric substituents are known in the art.The lipophilic substituent includes an acyl group, a sulphonyl group, anN atom, an O atom or an S atom which forms part of the ester, sulphonylester, thioester, amide or sulphonamide. The lipophilic moiety mayinclude a hydrocarbon chain having 4 to 30 C atoms, preferably between 8and 12 C atoms. It may be linear or branched, saturated or unsaturated.The hydrocarbon chain may be further substituted. It may be cycloalkaneor heterocycloalkane. The peptide may be modified at the N-terminal,C-terminal or both. The polymer or compound (binding partner) ispreferably linked to an amino, carboxyl or thio group and may be linkedby N-termini or C-termini of side chains of any amino acid residue. Thepolymer or compound (binding partner) may be conjugated to the sidechain of any suitable residue. The polymer or compound (binding partner)may be conjugated via a spacer. The spacer may be a natural or unnaturalamino acid, succinic acid, lysyl, glutamyl, asparagyl, glycyl,beta-alanyl, gamma-amino butanoyl. The polymer or compound (bindingpartner) may be conjugated via an ester, a sulphonyl ester, a thioester,an amide, a carbamate, a urea, a sulphonamide. A person skilled in theart is aware of suitable means to prepare the described conjugate.

“Fragment” means a segment of a protein selected from SEQ ID NO's: 1 to14, the fragment typically being 7 to 37 contiguous amino acids inlength, and generally having a charge of between −9 and +3; a c-terminalamino acid that typically is not cysteine (C) or methionine (M); and ann-terminal amino acid that typically is not cysteine (C), histidine (H),proline (P) or threonine (T). The charge of a peptide, fragment orregion is determined using the method of Cameselle, J. C., Ribeiro, J.M., and Sillero, A. (1986). Derivation and use of a formula to calculatethe net charge of acid-base compounds. Its application to amino acids,proteins and nucleotides. Biochem. Educ. 14, 131-136.

The term “natural” as applied to a peptide means a peptide that includes(a) a fragment of a plant protein, typically rice or pea protein, orvariants of pea protein including lentil, sweet pea, or chick pea orvariants of rice protein including oat, grass, corn, wild rice andbananas, or (b) a variant of the fragment of a plant protein, forexample a fragment of a homolog of the plant protein. The peptides orfragments of the invention may be isolated from plant proteins or madesynthetically using methods known to a person skilled in the art anddescribed herein.

“C-terminal domain” as applied to a fragment means the first three aminoacids at the c-terminus of the fragment.

“N-terminal domain” as applied to a fragment means the last three aminoacids at the n-terminus of the fragment.

“Bioactive” as applied to a peptide or fragment means having a healthpromoting effect when administered a mammal, for example one or more ofglucose transport promoting, anti-bacterial, anti-inflammatory, orcellular growth or proliferation promoting. In one embodiment, the term“bioactive” means cellular growth promoting.

“Growth promoting” or “growth promoting activity” as applied to apeptide or fragment means a peptide or fragment that is capable ofincreasing elastin production or cellular proliferation of human skintreated with a 20 μM solution of peptide or fragment as described in theassay below.

“Glucose transport promoting” or “glucose transport promoting activity”as applied to a peptide or variant or fragment means a peptide, variantor fragment that is capable of increasing GLUT4 translocation intoskeletal muscle compared with an untreated control when employed at aconcentration of 2 μM in the following in-vitro assay. L6-GLUT4myc cellswere grown in 10% FBS and 2 μg/ml blasticidin. Cells were grown for48-72 hours before being seeded in 24-well plates at 15,000 cells perwell in 2% FBS and allowed to differentiate for 6 to 8 days prior toexperimentation. L6-GLUT4myc cells were serum-starved for three hoursprior to incubation with 100 nM of insulin for 30 mins, or 200, 20, 2.0and 0.2 μM of SP, and 2, 1, 0.5 and 0.25 mg/ml of peptide/peptidecomposition for 3 hours respectively. A 3-hour incubation period wasselected based on previous findings identifying that incubation withbranch chain amino acid containing di-peptides for 3 hours increasesglucose uptake in L6 myotubes 1. Treatments were staggered in order todetermine GLUT4myc translocation at the same time point. The quantity ofmyc-tagged GLUT4 at the cell surface was measured by antibody-coupledcolorimetric assay. Briefly, after incubation with either insulin for 30mins or synthetic peptide or peptide composition for 3 hoursrespectively, L6-GLUT4myc cells were fixed via incubation with 3%paraformaldehyde (PFA). A 0.1 M glycine solution was then added toquench PFA and cells were blocked with 5% goat serum. The myotubemonolayer was exposed to anti-myc antibody and then incubated withperoxidase conjugated donkey anti-mouse IgG. 1 mL of o-phenylenediaminedihydrochloride (OPD) reagent was added to each well and this reactionwas stopped by adding 250 μl/well of 3 M HCL. To determine GLUT4translocation to cell surface, a measured aliquot of each condition wasdetermined spectrophotometrically on a plate reader using absorbance at492 nm. Preferably the peptide or fragment is capable of increasingGLUT4 translocation compared with an untreated control by at least 50%(i.e a relative unit increase in GLUT4 translocation of 1% to 1.5%).

“Antibacterial” or “antibacterial activity” as applied to a peptide orfragment means a peptide or fragment that is capable of visiblyinhibiting the growth of a bacteria in the following agar-plate basedgrowth inhibition assay: Peptide stock=5 mg/mL dissolved in DMSO.Bacterial inoculums were adjusted to McFarland 0.5 standard and MHAplates swabbed. Blank disks were placed in the plates and 10 μL of eachcompound (at 64 μg/mL—maximum concentration tested) added. Plates wereincubated at 37° C. for 16-18 hours. Appropriate controls (DMSO;Mueller-Hinton media alone; and two antibiotic discs—ciprofloxacin andtetracycline) were also performed.

“Anti-inflammatory” as applied to a peptide or fragment means a peptideor fragment that is capable of significantly reducing the secretion ofTNFα by LPS-stimulated J774.2 macrophages (compared with untreatedLPS-stimulated J774.2 macrophages) when the macrophages are treated with100 μM of the peptide or fragment. J774.2 macrophages were treated with100 μM of synthetic peptide for 24 hours and then stimulated with (A)LPS (10 ng/ml) for five hours or (B) LPS (10 ng/ml) for 5 hours followedby ATP (5 mM) for one hour. Supernatant was collected and levels of TNFαwere determined by ELISA.

“Enriched in peptides having a molecular weight of less than 10 KD” asapplied to a composition of the invention means that the dry weight % ofpeptides in the composition having a molecular weight of less than 10 KDis greater than the dry weight % of polypeptide/protein in thecomposition having a molecular weight of 10 KD or greater.

“Homolog” of a reference protein should be understood to mean a proteinfrom a different species of plant having at least 60%, 70%, 80%, 90%,91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence homology with thereference protein. Thus, for example, homologs of pea protein P13918include:

>gi|137584|sp|P08438.1|VCL_VICFA RecName: Full=Vicilin; Flags: Precursor[Vicia faba] >gi|22057|emb|CAA68559.1| vicilin [Vicia faba var.minor] >gi|383931031|gb|AFH56916.1| vicilin [Viciafaba] >gi|502105533|ref|XP_004492829.1| PREDICTED: vicilin-like isoformX1 [Cicer arietinum] ChickPea >gi|29539109|emb|CAD87730.1| allergen Lenc 1.0101 [Lens culinaris] Lentil

A “variant” of a growth promoting fragment shall be taken to mean afragment having an amino acid sequence that is substantially identicalto the reference growth promoting fragment, and which has growthpromoting activity as defined above. Thus, for example, the term shouldbe taken to include fragments that are altered in respect of one or moreamino acid residues. Preferably such alterations involve the insertion,addition, deletion and/or substitution of 5 or fewer amino acids, morepreferably of 4 or fewer, even more preferably of 3 or fewer, mostpreferably of 1 or 2 amino acids only. Insertion, addition andsubstitution with natural and modified amino acids is envisaged. Thevariant may have conservative amino acid changes, wherein the amino acidbeing introduced is similar structurally, chemically, or functionally tothat being substituted. Generally, the variant will have at least 70%amino acid sequence homology, preferably at least 80% sequence homology,more preferably at least 90% sequence homology, and ideally at least95%, 96%, 97%, 98% or 99% sequence homology with the reference growthpromoting fragment.

In this specification, the term “SEQ IDentity” should be understand tocomprise both SEQ IDentity and similarity, i.e. a variant (or homolog)that shares 70% SEQ IDentity with a reference sequence is one in whichany 70% of aligned residues of the variant (or homolog) are identical toor conservative substitutions of the corresponding residues in thereference sequence across the entire length of the sequence. Sequenceidentity is the amount of characters which match exactly between twodifferent sequences. Hereby, gaps are not counted and the measurement isrelational to the shorter of the two sequences. In terms of “sequencehomology”, the term should be understood to mean that a variant (orhomolog) which shares a defined percent similarity or identity with areference sequence when the percentage of aligned residues of thevariant (or homolog) are either identical to, or conservativesubstitutions of, the corresponding residues in the reference sequenceand where the variant (or homolog) shares the same function as thereference sequence. This alignment and the percent homology or sequenceidentity can be determined using software programs known in the art, forexample, one alignment program is BLAST, using default parameters.

Details of these programs can be found at the following Internetaddress: <www.ncbi.nlm.nih.gov/blast/Blast.cgi>.

Variants of SEQUENCE ID NO: 448 (QSFLLSGNQ)

Variants of SEQ ID NO: 448 (QSFLLSGNQ) including variants having 1 or 2conservative amino acid substitutions, 1, 2 to 3 non-conservative aminoacid substitutions, 1-2 amino acid additions, 1, 2 or 3 amino aciddeletions, are provided below:

One Conservative Amino Acid Substitution:

(SEQ ID NO'S 418-421) QSFILSGNE, ESFLLSGNQ, QSYLLSGNQ, QSFLLSGDQ

Two Conservative Amino Acid Substitutions:

(SEQ ID NO'S 422 to 426) QSYLLSGNE, ESFLLSGNE, ESYLLSGNQ, QSFLLSGDE,QSYLLSGDQ

One Non-Conservative Amino Acid Substitution

(SEQ ID NO'S 427 to 431) QSFRLSGNQ, QSFLLSYNQ, QFFLLSGNQ, QSFLLSGAQ,QSFLLSGNP

Two Non-Conservative Amino Acid Substitution

(SEQ ID NO'S 432 to 436) QSFRRSGNQ, QSFLLSYIQ, QFFLLSGNL, QSFLLSGAQ,QSFLLSGNP

One or Two Amino Acid Additions

(SEQ ID NO'S 437 to 441) QSFLLSGNQQ, QSFLLLSGNQ, AQSFGLLSGNQ,RQSFLLISGNQ, QSFLLSGNQK

One, Two or Three Amino Acid Deletions

(SEQ ID NO'S 442 to 451) QFLLSGNQ, SFLLSGNQ, QSFLLSGN, QSFLLGNQ,QSFLSGNQ, QSLLSGNQ, SFLLSGNQ, QSFLLSGN, SFLLSGN, QSFSGNQ

The term “variant” also includes fragment of peptides of the invention.“Fragment of a peptide of the invention” or “peptide fragment” means afragment of one of the peptides of the invention having at least 3, 4,5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 or 22amino acids and that typically has a bioactivity, for exampleanti-inflammatory activity, anti-ageing activity, glucose transportpromoting activity, or anti-bacterial activity. In one embodiment, thefragment consists of at least 30%, 40%, 50%, 60%, 70%, 80%, or 90% ofthe reference sequence. Thus, the invention also provides bioactivefragments of the peptides of the invention, and peptides comprising oneor more of these fragments. In one embodiment, the fragments arebioactive. In one embodiment, the fragments are cellular growth orproliferation promoting fragments. Examples of fragments of peptides ofthe invention are provide in SEQ ID NO'S 494 to 524.

“Anti-ageing” means inhibiting or slowing the appearance of ageing of ahuman skin and/or reversing the appearance of ageing. “Slowing orinhibiting ageing of the skin” means slowing or inhibiting the ageingprocess in the skin, and/or reversing the appearance of ageing.

“Disease or condition characterised by damaged dermal or epithelialcells or tissue” means any condition or disease that results in damageddermal or epithelial tissue or cells or organs. One example is traumawhich often results in damaged skin. Another example is an inflammatoryskin condition such as psoriasis or eczema which often results indamaged skin. Another example is an inflammatory disorder of the lowerintestines which can result in damaged epithelial cells/tissue liningthe lower intestines. Another example is damaged epithelial cells/tissuelining the lower intestines caused by ingestion of a toxic or damagingsubstance, for example toxic chemicals or drugs. Another example iscancer, for example bowel cancer, which can result in damaged epithelialtissue in the bowel. Another condition is a peripheral inflammatorydisorder such as atopic dermatitis which can result in damage to theskin in humans.

“Inflammatory disorder” means an immune-mediated inflammatory conditionthat affects humans and is generally characterised by dysregulatedexpression of one or more cytokines. Examples of inflammatory disordersinclude skin inflammatory disorders, inflammatory disorders of thejoints, inflammatory disorders of the cardiovascular system, certainautoimmune diseases, lung and airway inflammatory disorders, intestinalinflammatory disorders. Examples of skin inflammatory disorders includedermatitis, for example atopic dermatitis and contact dermatitis, acnevulgaris, and psoriasis. Examples of inflammatory disorders of thejoints include rheumatoid arthritis. Examples of inflammatory disordersof the cardiovascular system are cardiovascular disease andatherosclerosis. Examples of autoimmune diseases include Type 1diabetes, Graves disease, Guillain-Barre disease, Lupus, Psoriaticarthritis, and Ulcerative colitis. Examples of lung and airwayinflammatory disorders include asthma, cystic fibrosis, COPD, emphysema,and acute respiratory distress syndrome. Examples of intestinalinflammatory disorders include colitis and inflammatory bowel disease.Other inflammatory disorders include cancer, hay fever, periodontitis,allergies, hypersensitivity, ischemia, depression, systemic diseases,post infection inflammation and bronchitis. The invention also relatesto a peptide or composition of the invention for use in treating aninflammatory disorder in a mammal.

“Metabolic disorder” should be understood to include pre-diabetes,diabetes; Type-1 diabetes; Type-2 diabetes; metabolic syndrome; obesity;diabetic dyslipidemia; hyperlipidemia; hypertension;hypertriglyceridemia; hyperfattyacidemia; hypercholerterolemia;hyperinsulinemia, and MODY. The invention also relates to a peptide orcomposition of the invention for use in treating a metabolic disorder ina mammal.

“Disease or condition characterised by damaged dermal or epithelialcells or tissue” means any condition or disease that results in damageddermal or epithelial tissue or cells or organs. One example is traumawhich often results in damaged skin. Another example is an inflammatoryskin condition such as psoriasis or eczema which often results indamaged skin. Another example is an inflammatory disorder of the lowerintestines which can result in damaged epithelial cells/tissue liningthe lower intestines. Another example is damaged epithelial cells/tissuelining the lower intestines caused by ingestion of a toxic or damagingsubstance, for example toxic chemicals or drugs. Another example iscancer, for example bowel cancer, which can result in damaged epithelialtissue in the bowel. Another condition is a peripheral inflammatorydisorder such as atopic dermatitis which can result in damage to theskin in humans.

“Disease or condition characterised by bacterial infection” means anycondition or disease characterised having a pathology caused by growthof bacteria or by bacterial infection, including for example MRSA,salmonella, listeria, bacterial pneumonia, Staphylococcal foodpoisoning, bacterial meningitis. Specific examples are provided in<en.wikipedia.org/wiki/List_of_infectious_diseases>.

“Man-made” as applied to comestible products should be understood tomean made by a human being and not existing in nature.

“Maintaining or restoring gut health” means reducing and/or regulatingthe pro-inflammatory response in the gut and more specifically theepithelial cells. The healthy microbiome offers some protection againstpathogenic viruses and bacteria, and their presence is needed to guidethe development of our immune system. It has been shown that thesebacteria can react to human signals of stress, sickness, or age whichcan be manifested by inflammation and as a consequence switch on theirvirulence genes and cause or contribute to disease. Having the abilityto reduce and maintain at healthy levels the inflammatory response canhelp maintain the healthy bacteria. Digestive problems, which comprisethe number one health problem in North America, appear to be occurringwith more frequency in recent years. One way to maintain digestivehealth is to maintain proper inflammation and intestinal flora.

“Improving muscle status” means improving the muscle health, for examplepromoting skeletal muscle protein synthesis, skeletal glucoseabsorbtion, improving lean tissue mass in therapeutic or non-therapeuticcontext, promoting muscle recovery generally after activity exercise, orimproving muscle performance. The methods or uses may be therapeutic ornon-therapeutic. The term “improving lean tissue mass status” should beunderstood to mean increasing lean tissue mass, or inhibiting orpreventing the rate of lean tissue mass degradation.

“Promoting muscle recovery” means causing an increase in absorbtion ofglucose in skeletal muscle compared with untreated skeletal muscle.

“Disease or condition characterised by lethargy or low energy levels”means any condition or disease characterised by a feeling or tirednessor low energy. Examples include allergies, asthma, anaemia, cancer andits treatments, chronic pain, heart disease, infection, depression,eating disorders, grief, sleeping disorders, thyroid problems,medication side effects, alcohol use, or drug use.

“Maintaining or restoring muscle health” means helping retain or restoremammalian muscle health resulting from damage incurred during exercise.By promoting glucose transport in skeletal muscle the peptides promoterecovery from exercise, and relieve muscle soreness/pain and injuryconnected with exercise. They can also be used to decrease and preventmuscle cramping, and to allow a faster recovery from muscle cramping.Cramping can result from physical stress, mental stress, and orRepetitive Strain Injury stress. By promoting glucose transport thepeptides help reduce Myopathy of the muscle, and help prevent Sarcopeniain mammals, promote recovery from injuries during exercise, and relievemuscle soreness/pain and injury connected with exercise. The inventionalso relates to a peptide or composition of the invention for use inmaintaining or restoring muscle health in a mammal.

In this specification, the term “substantially all” as applied to a listof peptides should be understood to mean at least 60%, 70%, 80%, 90% or95% of the peptides.

“Man-made” as applied to comestible products should be understood tomean made by a human being and not existing in nature.

BRIEF DESCRIPTION OF THE FIGURES

FIGS. 1 to 100: Effect of synthetic peptides of the invention onproliferation of Human Dermal Fibroblasts (HDF).

FIG. 1: peptide of SEQ ID NO: 121,

FIG. 2: peptide of SEQ ID NO: 105,

FIG. 3: peptide of SEQ ID NO: 249,

FIG. 4: peptide of SEQ ID NO: 226,

FIG. 5: peptide of SEQ ID NO: 84,

FIG. 6: peptide of SEQ ID NO: 330,

FIG. 7: peptide of SEQ ID NO: 181,

FIG. 8: peptide of SEQ ID NO: 83,

FIG. 9: peptide of SEQ ID NO: 247,

FIG. 10: peptide of SEQ ID NO: 97,

FIG. 11: peptide of SEQ ID NO: 74,

FIG. 13: peptide of SEQ ID NO: 168,

FIG. 14: peptide of SEQ ID NO: 151,

FIG. 15: peptide of SEQ ID NO: 470,

FIG. 16: peptide of SEQ ID NO: 257,

FIG. 17: peptide of SEQ ID NO: 258,

FIG. 18: peptide of SEQ ID NO: 457,

FIG. 19: peptide of SEQ ID NO: 499,

FIG. 20: peptide of SEQ ID NO: 253,

FIG. 21: peptide of SEQ ID NO: 222,

FIG. 22: peptide of SEQ ID NO: 272,

FIG. 23: peptide of SEQ ID NO: 252,

FIG. 24: peptide of SEQ ID NO: 248,

FIG. 25: peptide of SEQ ID NO: 472,

FIG. 26: peptide of SEQ ID NO: 365,

FIG. 27: peptide of SEQ ID NO: 502,

FIG. 28: peptide of SEQ ID NO: 496,

FIG. 29: peptide of SEQ ID NO: 98,

FIG. 30: peptide of SEQ ID NO: 454,

FIG. 31: peptide of SEQ ID NO: 85,

FIG. 32: peptide of SEQ ID NO: 453,

FIG. 33: peptide of SEQ ID NO: 158,

FIG. 34: peptide of SEQ ID NO: 464,

FIG. 35: peptide of SEQ ID NO: 73,

FIG. 36: peptide of SEQ ID NO: 359,

FIG. 37: peptide of SEQ ID NO: 124,

FIG. 38: peptide of SEQ ID NO: 112,

FIG. 39: peptide of SEQ ID NO: 733,

FIG. 40: peptide of SEQ ID NO: 728,

FIG. 41: peptide of SEQ ID NO: 727,

FIG. 42: peptide of SEQ ID NO: 730,

FIG. 43: peptide of SEQ ID NO: 731,

FIG. 44: peptide of SEQ ID NO: 148,

FIG. 45: peptide of SEQ ID NO: 343,

FIG. 46: peptide of SEQ ID NO: 345,

FIG. 47: peptide of SEQ ID NO: 484,

FIG. 48: peptide of SEQ ID NO: 729,

FIG. 49: peptide of SEQ ID NO: 456,

FIG. 50: peptide of SEQ ID NO: 494,

FIG. 51: peptide of SEQ ID NO: 723,

FIG. 52: peptide of SEQ ID NO: 722,

FIG. 53: peptide of SEQ ID NO: 498,

FIG. 54: peptide of SEQ ID NO: 475,

FIG. 55: peptide of SEQ ID NO: 718,

FIG. 56: peptide of SEQ ID NO: 337,

FIG. 57: peptide of SEQ ID NO: 500,

FIG. 58: peptide of SEQ ID NO: 717,

FIG. 59: peptide of SEQ ID NO: 297,

FIG. 60: peptide of SEQ ID NO: 340,

FIG. 61: peptide of SEQ ID NO: 719,

FIG. 62: peptide of SEQ ID NO: 726,

FIG. 63: peptide of SEQ ID NO: 725,

FIG. 64: peptide of SEQ ID NO: 724,

FIG. 65: peptide of SEQ ID NO: 720,

FIG. 66: peptide of SEQ ID NO: 721,

FIG. 67: peptide of SEQ ID NO: 503,

FIG. 68: peptide of SEQ ID NO: 474,

FIG. 69: peptide of SEQ ID NO: 504,

FIG. 70: peptide of SEQ ID NO: 114,

FIG. 71: peptide of SEQ ID NO: 505,

FIG. 72: peptide of SEQ ID NO: 482,

FIG. 73: peptide of SEQ ID NO: 479,

FIG. 74: peptide of SEQ ID NO: 477,

FIG. 75: peptide of SEQ ID NO: 410,

FIG. 76: peptide of SEQ ID NO: 475,

FIG. 77: peptide of SEQ ID NO: 497,

FIG. 78: peptide of SEQ ID NO: 480,

FIG. 79: peptide of SEQ ID NO: 463,

FIG. 80: peptide of SEQ ID NO: 465,

FIG. 81: peptide of SEQ ID NO: 467,

FIG. 82: peptide of SEQ ID NO: 461,

FIG. 83: peptide of SEQ ID NO: 341,

FIG. 84: peptide of SEQ ID NO: 468,

FIG. 85: peptide of SEQ ID NO: 285,

FIG. 86: peptide of SEQ ID NO: 496,

FIG. 87: peptide of SEQ ID NO: 146,

FIG. 88: peptide of SEQ ID NO: 478,

FIG. 89: peptide of SEQ ID NO: 452,

FIG. 90: peptide of SEQ ID NO: 495,

FIG. 91: peptide of SEQ ID NO: 403,

FIG. 92: peptide of SEQ ID NO: 455,

FIG. 93: peptide of SEQ ID NO: 270,

FIG. 94: peptide of SEQ ID NO: 501,

FIG. 95: peptide of SEQ ID NO: 473,

FIG. 97: peptide of SEQ ID NO: 471,

FIG. 98: peptide of SEQ ID NO: 460,

FIG. 99: peptide of SEQ ID NO: 93, and

FIG. 100: peptide of SEQ ID NO: 462.

FIG. 101: Effect of synthetic peptide of the invention (SEQ ID NO: 42)on elastin synthesis of Human Dermal Fibroblasts (HDF).

FIG. 102: Effect of synthetic peptide of the invention (SEQ ID NO: 42)on collagen synthesis of Human Dermal Fibroblasts (HDF).

FIG. 103: Effect of synthetic peptide of the invention (SEQ ID NO: 701)on elastin synthesis of Human Dermal Fibroblasts (HDF).

FIG. 104: Effect of synthetic peptide of the invention (SEQ ID NO: 701)on collagen synthesis of Human Dermal Fibroblasts (HDF).

FIG. 105: Effect of synthetic peptide of the invention (SEQ ID NO: 246)on elastin synthesis of Human Dermal Fibroblasts (HDF).

FIG. 106: Effect of synthetic peptide of the invention (SEQ ID NO: 246)on collagen synthesis of Human Dermal Fibroblasts (HDF).

FIG. 107: Effect of synthetic peptide of the invention (SEQ ID NO: 284)on elastin synthesis of Human Dermal Fibroblasts (HDF).

FIG. 108: Effect of synthetic peptide of the invention (SEQ ID NO: 245)on elastin synthesis of Human Dermal Fibroblasts (HDF).

FIG. 109: Effect of synthetic peptide of the invention (SEQ ID NO: 245)on collagen synthesis of Human Dermal Fibroblasts (HDF).

FIG. 110. shows the integrity controls and viability controls for theassay system.

FIG. 111. % of elastin expression in superficial dermis compared tocontrol (water or DMSO) for peptides P1, P2 and P3

* shows significant increases of elastin expression in superficial ANDmiddle dermis.

FIG. 112. % of elastin expression in middle dermis compared to control(water or DMSO) for peptides P1, P2 and P3.

* shows significant increases of elastin expression in superficial ANDmiddle dermis.

FIG. 113. % of cell proliferation in the basal layer of epidermiscompared to control (water or DMSO) for peptides P6 and P8, and peptidecompositions P9 and P10

* shows significant increases.

FIG. 114. Histological analysis of the elastic fibers (+catechin, ×200)

FIG. 115. Immunohistochemical evaluation of the mitotic index (Ki67,×400)

DETAILED DESCRIPTION OF THE INVENTION Example 1—Cell Proliferation Assay

BrDu is incorporated into newly synthesised DNA strands of activelyproliferating cells. Following partial denaturation of double strandedDNA, Brdu is detected immunochemically allowing the assessment of thepopulation of cells which are synthesizing DNA.

Human Dermal Fibroblasts (HDF—Sigma 10605a) were seeded in a 96 wellplate at 10,000 cells per well in DMEM containing 10% fetal calf serum(FCS), 1% Pen/strep, 1% L-glutamine and allowed to adhere for 24 h.

Following the initial 24 h incubation the cells were incubated with 5μg/ml, 0.5 μg/ml or 0.05 μg/ml synthetic peptide for 24 h respectively.

After 18 h incubation with synthetic peptides 20 μl BrDu reagent wasadded to each well.

At 24 h incubation the cell were fixed and the amount of 2-DG6P wasmeasured using the BrdU Cell Proliferation Assay, all steps were carriedout according to the manufacturer's instructions.

Results were calculated as a percentage of the untreated control. Anincrease in optical density reading indicates greatER incorporation ofBrDu and increase cell proliferation.

The results are shown in FIGS. 1-100 and Table 1 below.

TABLE 1 FIG. NO SEQ ID INCREASE IN PROLIFERATION 1 121 48% 2 105 40% 3249 30% 4 226 30% 5 84 20% 6 330 18% 7 181 33% 8 83 32% 9 247 28% 10 9726% 11 74 29% 12 13 168 14 151 15 470 119%  16 257 118%  17 256 117%  18457 114%  19 499 113%  20 253 112%  21 222 110%  22 272 97% 23 252 111% 24 248 86% 25 472 77% 26 365 58% 27 502 68% 28 496 51% 29 98 49% 30 45438% 31 85 35% 32 453 25% 33 158 21% 34 464 18% 35 73 16% 36 359 15% 37124 15% 38 112 15% 39 733 40 728 41 727 42 730 43 731 44 148 45 343 46345 47 484 48 729 49 456 50 494 51 723 52 722 53 498 54 475 13% 55 71856 337  8% 57 500  6% 58 717 59 297 60 340 61 719 62 726 63 725 64 72465 720 66 721 67 503 125%  68 474 121%  69 504 119%  70 114 119%  71 505118%  72 482 113%  73 479 106%  74 477 81% 75 410 73% 76 475 69% 77 49758% 78 480 102%  79 463 100%  80 465 96% 81 467 90% 82 461 85% 83 34183% 84 468 82% 85 285 81% 86 496 81% 87 146 80% 88 478 76% 89 452 76% 90495 68% 91 403 51% 92 455 47% 93 270 47% 94 501 43% 95 473 41% 96 39% 97471 38% 98 460 38% 99 93 26% 100 462 15%

Example 2—Collagen Production Assay

Hydroxyproline in tissue preparations is a direct measure of the amountof collagen present. FIRELISA Human Hydroxyproline ELISA kit assay isdesigned to measure hydroxyproline in tissue or peptide compositions.

Human Dermal Fibroblasts (HDF Sigma 10605a) were seeded in 24 wellplates at 50,000 cells per well in DMEM containing 10% fetal calf serum(FCS), 1% Pen/strep, 1% L-glutamine and allowed to adhere for 24 h.

Following the initial 24 h incubation the cells were incubated with 5μg/ml, 1 μg/ml or 0.1 μg/ml synthetic peptide for 96 h respectively.

After treatment the cells were lysed using 4 freeze thaw cycles inliquid nitrogen. The lysed cells were centrifuged and 50 μl/ml of eachsupernatant was assayed using the FIRELISA Human Hydroxyproline ELISAkit. All steps were carried out according to the manufacturer'sinstructions.

Results were calculated as a percentage of the untreated control. Anincrease in optical density reading indicates an increase collagencontent.

The results are shown in FIGS. 102, 104, 106 and 109

Example 3—Elastin Production Assay

Elastin is a highly elastic protein in connective tissue and allows manytissues in the body to resume their shape after stretching orcontracting. FIRELISA Human Elastin ELISA kit assay is designed tomeasure Elastin in tissue or protein/peptide compositions.

Human Dermal Fibroblasts (HDF) were seeded in 24 well plates at 50,000cells per well in DMEM containing 10% fetal calf serum (FCS), 1%Pen/strep, 1% L-glutamine and allowed to adhere for 24 h.

Following the initial 24 h incubation the cells were incubated with 5μg/ml, 1 μg/ml or 0.1 μg/ml synthetic peptide for 96 h respectively.

After treatment the cells were lysed using 4 freeze thaw cycles inliquid nitrogen. The lysed cells were centrifuged and 50 μl/ml of eachsupernatant was assayed using the FIRELISA Human Elastin ELISA kit. Allsteps were carried out according to the manufacturer's instructions.

Results were calculated as a percentage of the untreated control. Anincrease in optical density reading indicates an increase collagencontent.

The results are shown in FIGS. 101, 103, 105, 107, 108 and 109.

Example 4—Elastin and Cell Proliferation Assays

TABLE 2 Test items. Orange bands correspond to samples dissolved intoDMSO 0.3% instead of water. Intertek Item Denomination ConcentrationProvider Nature reference Solubility Storage Peptide 1 E_280_PJ  20 μMNuritas Peptide 14-CHL- Water −80° C. 0723-01 Peptide 2 I_222two_IN  20μM Nuritas Peptide 14-CHL- Water Ambient 0723-02 Peptide 3 E_134_two_IN 20 μM Nuritas Peptide 14-CHL- Water −80° C. 0723-03 Peptide 4E_30two_IN  20 μM Nuritas Peptide 14-CHL- Water −80° C. 0723-04 Peptide5 E_121two_IN  20 μM Nuritas Peptide 14-CHL- Water −80° C. 0723-05Peptide 6 I_10two_IN  20 μM Nuritas Peptide 14-CHL- DMSO −80° C. 0723-060.3% Peptide 7 I_41two_IN  20 μM Nuritas Peptide 14-CHL- DMSO −80° C.0723-07 0.3% Peptide 8 E_41_PJ  10 μM* Nuritas Peptide 14-CHL- Water−80° C. 0723-08 Composition E_2_IN 500 μg/mL Nuritas Composition 14-CHL-Water −80° C. P9 of peptides 0723-09 Composition I_2_IN 500 μg/mLNuritas Composition 14-CHL- Water −80° C. P10 of peptides 0723-10

Equipment

Incubator, Flow Laminar Chamber, Sterile Polished Plastic Rod, Pipettor,Maintenance medium, Plate 6 well, Plate 24 well.

Reagents

MTT, PBS, SDS, Formaldehyde, Xylene, Ethanol absolute, Dulbecco'sphosphate-buffered saline (DPBS), Metal Enhanced DAB substrate kit, ABCperoxidase staining kit, Citric acid, Sodium hydroxide 2N, Hydrogenperoxide 30%, Anti-Filaggrin, Anti-rabbit IgG-Biotin, Tween 20.

Test System

Nature: Human skin tissue 5 mm diameter

Batch number: EXP004050B009 and EXP004050B011

Provider: Laboratoire Biopredic International—8-18 rue Jean Pecker—35000Rennes—

France. Tel: +33 (0)2.99.14.36.14—Fax: +33 (0)2.99.54.44.72.

Certificates of analysis are present in Annex 1.

Two batches are used for the assay. Batch EXP004050B005 is used forexperiment day 1, and Batch EXP004050B006 is used for experiment day 5.

Maintenance Medium

Maintenance Medium: Batch n°: MIL 218C

Provider: Laboratoire Biopredic International—8-18 rue Jean Pecker—35000Rennes—France.

Peptides Tested

P1: SEQ ID NO: 283 P2: SEQ ID NO: 246 P3: SEQ ID NO: 284(SEQ ID NO: 776) P4: RPYYSNAPQEIF (SEQ ID NO: 777) P5: VLLEQQEQEPQHP6: SEQ ID NO: 245 (SEQ ID NO: 778) P7: QQYGIAASPFLQSAAP8: SEQ ID NO: 42

Compositions Tested

P9 (14-CHL-0723-09) is the Pea composition (SEQ ID NO: 50, 85, 74, 140,82, 136, 189, 77, 169, 149, 171, 178, 143, 127, 190, 141, 147, 133, 186,125, 122, 119, 87, 90, 86, 89, 138, 129, 123, 120, 117, 113, 110, 121,105, 98, 55, 161, 19, 317, 135, 130, 146, 177, 160, 170, 188, 83, 78,36, 96, 159, 26, 330, 168, 148, 184, 151, 151, 165, 114, 284) P10(14-CHL-0723-010) is the Rice composition (SEQ ID Numbers: 245, 246,263, 250, 257, 259, 276, 255, 251, 264, 256, 266, 274, 270, 269, 356,245, 380, 262, 258, 356, 218, 252, 358, 271, 253, 344, 275, 272, 226,224, 220, 248, 261, 265, 373, 375, 247, 249, 363, 273, 343, 273, 362)

Application Method

Skin explants were prepared from abdominal plastic surgery. Someexplants were delipidated with alcohol to obtain a dehydrated skin.

These explants were maintained in maintenance medium supplied by theprovider Biopredic International for 5 days. Test items are appliedtwice per day with 5 μL per explant.

At the end of the test, viabilities controls are realized with the MTTon two explants, the third explant is fixed in the formaldehyde 4% forhistology and cell staining.

For each time of analysis (D1 and D5), histologies on delipidatedexplants, treated explants with test items, the DMSO 0.3% control andwater control, are performed.

After receipt in the laboratory, each skin explant in the maintenancemedium is delipidated with 5 μL alcohol during 3 hours.

After 3 hours, all skin explants are treated two per day with testitems, and they are incubated at 37° C.+/−2° C., 5% CO2 for 1 day or 5days.

Integrity of the system is realized at day 1 and day 5 with a viabilitycontrol with MTT.

Immunostaining

Histology is realized by the laboratory Gredeco and the immunostainingto elastin and Ki67 are realized by the same laboratory. Immunostainingto filaggrin is realized by the laboratory Intertek.

The detection of elastin (rabbit monoclonal antibody, clone P15502,LSBio) is performed using an immunoperoxidase technique two layers (ABCkit, Vector Laboratories) and revealed by AEC(3-amino-9-éthylcarbazole). The immunohistochemical staining intensityin the elastic fibers is evaluated using a semi-quantitativehistological score.

Epithelial proliferation was analyzed by immunohistochemistry usinganti-Ki67 antibody. Immunodetection was performed using an indirectimmunoperoxidase technique three layers, amplified (DAKO kit) andrevealed by AEC (3-Amino-9-ethylcarbazole). Counting the number oflabeled cells (keratinocytes of the basal layer of the epidermis) isperformed and provides the total number of basal cells to calculate the% of labeled cells.

The specific staining of filaggrin is performed with an immunoperoxidasestaining (ABC kit, Fisher). The intensity of immunohistochemical markerin the epidermis is evaluated relative to the negative control of thesolvent (Water or DMSO 0.3%).

C: Results

Viability Control

The integrity control and the viability control are present in FIG. 1.These controls do allow to validate the assay system. The viabilityis >50% for test items, and they do not show a cytotoxicity according tothe test.

Immunostaining

Elastin Expression

The elastic fibers of the dermis were revealed by staining with thecatechin and morphometrically quantified by analysis bycomputer-assisted image. The percentage area taken up by elastic fibersin the dermis was calculated in the dermis and the average superficialdermis. Results are presents in Table 1, FIG. 2 and FIG. 3.

TABLE 3 Morphometric quantification of elastic fibers in the superficialdermis (%) and middle dermis. Orange bands correspond to samplesdissolved into DMSO 0.3% instead of water. Morphometric Morphometricquantification of elastic quantification of elastic fibers in thesuperficial fibers in the middle dermis (%) dermis (%) Conditions D1 D5D1 D5 Dehydrated with 4.31 4.9 5 5.83 alcohol and hydrated with waterDehydrated 2.38 7.26 4.39 9.59 EGF (Epidermal 3.64 5.61 5.68 6.61 GrowthFactor) 10 ng/mL Dehydrated with 3.76 7.24 6 10.36 alcohol and hydratedwith DMSO 0.3% 0723.01 4.45 10.21 7.59 10.17 0723.02 6.09 7.59 11.759.08 0723.03 3 11.68 4.9 9 0723.04 3.28 8.94 5.22 9 0723.05 6.34 6.268.8 6.61 0723.06 3.8 4.03 4.54 8.67 0723.07 4 5.15 6.46 5.33 0723.08 2.75.32 3.52 7.27 0723.09 3.26 8.26 5.75 7.92 0723.10 4.1 8 5.73 8.34

Under the experimental conditions of the study, 0723-1 and 0723-3samples show an increase by twice of elastic fibers in the superficialdermis compared to control water (Figure), and an increase in the middledermis compared to the water control at D5.

The 0723-2 sample shows an increase doubled in the middle dermis at day1 compared to control water and an increase at day 5.

Ki67 Expression

The results of the immunohistochemical analysis of Ki67 are reported inTable and expressed as % of labelled at the basal layer of theepidermis. The Figure shows the percentage of Ki 67 cells compared tonegative controls (water or DMSO).

Immunohistochemical analysis of mitotic activity is shown in annex 4with a reminder of the average for each analysed conditions.

TABLE 4 % of Ki67 positive cells in the basal layer of the epidermis.Orange bands correspond to samples dissolved into DMSO 0.3% instead ofwater. Conditions D1 D5 Dehydrated with alcohol and hydrated with 19.093.53 water Dehydrated 17.05 1.76 EGF (Epidermal Growth Factor) 10 ng/mL25.11 4.2 Dehydrated with alcohol and hydrated with 17.2 2.61 DMSO 0.3%0723.01 18.57 3.92 0723.02 19.61 6.73 0723.03 22.01 10.04 0723.04 14.9711.36 0723.05 9.48 3.08 0723.06 31.97 5.04 0723.07 22.22 5.26 0723.0827.83 5.72 0723.09 31.02 2.4 0723.10 31.94 3.57

Under the experimental conditions of the study, test item 0723-06,0723-08, 0723-09 and 0723-010 show an increase in the number of mitoticcells compared to EGF at day 1. A decrease in the mitotic index wasobserved on day 5 compared to day 1 for all analysed conditions.

The decrease in this cell staining on day 5 is caused by the model.Indeed, after approximately 3 days cell turnover is exhausted on thismodel.

The invention is not limited to the embodiments hereinbefore describedwhich may be varied in construction and detail without departing fromthe spirit of the invention.

The invention claimed is:
 1. A composition formulated for topicalapplication to the skin of a human comprising a peptide selected fromSEQ ID NO: 284, SEQ ID NO: 285 and SEQ ID NO: 472, and a cosmeticallyacceptable excipient, wherein the composition is in the form of a gel,cream, lotion, ointment or emulsion.
 2. The composition of claim 1, inthe form of a gel, cream, lotion, or ointment.
 3. The composition ofclaim 1, comprising 0.1 to 5000 ppm of the peptide.
 4. The compositionof claim 1, comprising 0.5 to 500 ppm of the peptide.
 5. The compositionof claim 1, in the form of an emulsion.
 6. The composition of claim 1,wherein the cosmetically acceptable excipient is selected from adiluent, carrier, binder, lubricant, suspending agent, coating agent,preservative, stabilizers, dyes, vehicle, solubilizing agent, base,emollient, emulsifying agent, fragrance, humectant, or surfactants. 7.The composition of claim 1, wherein the cosmetically acceptableexcipient is selected from an emollient, emulsifying agent, orhumectant.
 8. A method of slowing aging of human skin comprisingtopically administering to the skin of a human a composition of claim 1.